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Biomedical subjects

W Xiao

Publications and source records attributed to W Xiao.

At least 127 records · Page 7Linked to original sources
PubMed1996

Mutagenicity and toxicity of the DNA alkylation carcinogens 1,2-dimethylhydrazine and azoxymethane in Escherichia coli and Salmonella typhimurium.

DNA alkylating agents such as 1,2-dimethylhydrazine (SDMH) and azoxymethane (AOM) are potent carcinogens and are widely used to induce colon tumors in experimental animals. However, standard bacterial mutagenesis assays have failed to detect the mutagenic effects of these chemicals. Using derivatives of a set of Escherichia coli test strains developed by Cupples and Miller (Proc. Natl. Acad. Sci. USA, 86, 5345, 1989), we hve demonstrated that under two conditions, SDMH and AOM induced point mutations by several-fold in a dose-dependent manner: (i) of six possible base substitutions, they only induced GC-->AT transitions; and (ii) the cells must be deficient in O6-methylguanine (O6MeG) DNA methyltransferase (MTase) activity. SDMH and AOM up to 200 microg/ml were unable to induce His+ revertants in a Salmonella Ames test strain TA1535 (GC-->AT); however, in the absence of mammalian S9 extract, His+ revertants increased up to 55-fold upon treatment of an isogenic Salmonella strain deficient in MTase activity. These results indicate that SDMH and AOM are indeed bacterial mutagens and that lesions induced by them are the target of DNA repair MTases, which probably include mutagenic and carcinogenic lesions such as O6MeG. Furthermore, variable responses of bacterial species to SDMH- and AOM-induced mutagenicity suggests a difference either in the metabolism of potential mutagens or in the repair of specific lesions. Since O6MeG is not only a mutagenic lesion but also a lethal lesion if left unrepaired, we compared the mutagenicity and toxicity of SDMH and AOM with an SN-type methylating carcinogen. N-methyl-N'-nitro-N-nitrosoguanidine, and conclude that SDMH and AOM are weak bacterial mutagens.

1,2-Dimethylhydrazine↗
PubMed1996

Neurobiological mechanisms of the meridian and the propagation of needle feeling along the meridian pathway.

The present experiments attempt to find the meridian phenomenon and how the needle feeling propagates along the given meridian channels. The neurobiological mechanisms of the meridian were studied with neuroelectrical recording from the motor neurons and CB-HRP retrograde histochemistry technique in both rats and cats. The results demonstrated that most, but not all, of alpha motor neurons supplying a muscle group of a given meridian were selectively activated by afferent inputs originating not only from homonymous or heterogeneous, but synergistic muscle, but also from the skin nerve overlying the muscle group of the homonymous meridian. However, the afferent inputs from the heterogeneous meridian have very weak or no effect. On the other hand, the labeled motor neurons supplying a given meridian muscles from a discrete longitudinal column with a definite bound in the lateral ventral horn. There are oriented dendro-dendristes projections between the labeled motor neurons. The characteristics of both selective responses of the motor neurons to afferent inputs and their neuro-anatomical arrangements in spinal cord offer neurobiological evidence for the meridian phenomenon.

Acupuncture Therapy↗
PubMed1996

[Analysis of DNA methylation pattern of c-myc oncogene in hepatocellular carcinoma].

OBJECTIVES: To assess whether hypomethylation of c-myc gene occours in HCC and to determine its clinical significance. METHODS: The methylation pattern at 3' end of c-myc gene was analyzed by Southern blot using methylation sensitive restrcition endonuclease enzymes Hpa I and Msp I in six normal liver tissues and 18 HCCs. RESULTS: At least 5 CCGG sites were methylated at 3' end of c-myc gene in 6 normal liver tissues. However, the normal hypermethylation pattern was absent in 11 hepatocellular carcinoms (HCC) which showed different degree of hypomethylation even if among subclonal cells of same tumour tissues. The hypomethylation was observed more often in markedly advanced HCC with metastasis and poorly differentiated histological grade. CONCLUSION: Scattered CpG at 3' end of c-myc gene was methylated in normal liver tissues. Methyl groups were deleted in different extent in HCC, which can lead to abnormal expression of c-myc gene. The hypomethylation of c-myc can reflect the malignlant extent of HCC and may serve as a useful prognostic indicator in clinical practice.

Carcinoma, Hepatocellular↗
PubMed1996

[Studies of the characteristic features of Ki-1 positive non-Hodgkin's lymphoma].

The clinical histopathological and immunophenotypic features in 5 patients with Ki-1 positive non-Hodgkin's lymphoma (NHL) were studied. When first seen, 4 patients presented enlargement of superficial lymph nodes, with skin lesions in 2 patients. Two patients in stage IV with fever, hepato-splenomegaly and bone marrow invasion, died. Histologically, the tumor cells showed diffused or patchy hyperplasia. The cells were relatively large in size, rich in bosophilic or slightly eosinophilic cytoplasm with irregularly-shaped nuclei, prominent nucleoli, and distinct anaplasia and pleomorphism. Some of the cells looked very much like the Reed-Sternberg cells. Multinucleated giant cells were seen. Immunophenotypically, all the cells were CD30 (Ki-1) and CD25 (IL-2 receptor) positive but CD15 (Leu M1) negative. Thus, the 5 patient T Ki-1 positive NHL were all of T cell type.

Adolescent↗
PubMed1996

[Changes of peripheral blood stem cells of patients with malignant hematopoietic tumor after chemotherapy].

To investigate the most appropriate condition for collecting stem cells during peripheral blood stem cells transplantation (PBSCT), we used colony forming unit-granulocyte, macrophage (CFU-GM) as index to observe the change of PBSC after chemotherapy in 12 patients with malignant hematopoietic tumor. The results showed that it was more suitable to collect PBSC when the number of leucocytes was 2 x 10(9)/L, neutrophils 0.8 x 10(9)/L and platelets 110 x 10(9)/L after chemotherapy for 2 weeks. After chemotherapy, the number of leucocytes were suppressed evidently. On the contrary, a lot of CFU-GM appeared during the recovery phase of peripheral hemogram. At the nadir of peripheral hemogram, the number of leucocytes correlated negatively with the number of CFU-GM (P < 0.05). The longer the time of chemotherapy, the less the CFU-GM that in the same patients. After chemotherapy the CD34+ cells which appeared early might not form CFU-GM, but the CD34+ cells at the recovery phase of peripheral hemogram had the same trend of change with CFU-GM.

Adolescent↗
PubMed1996

Essential region for self-replication of Coryneform bacteria plasmid pXZ10145.

A pTSK series of recombinant plasmids were constructed by cloning DNA fragments of pXZ10145 or its deleted deriviate pATN65 into plasmid vector pACYC177 of E. coli. Experiment results of Coryneform bacteria transformation with these pTSK plasmids allowed us to localize the essential region for self-replication on plasmid pXZ10145. The minimal replication region of the pXZ10145 was located on a 1.2kb Nael-Nrul DNA fragment in which only one open reading frame was found. This ORF was believed to be encoded a trans-acting replication factor. The replication origin (oriV) was locate on a 0.3kb NaeI-SalI fragment which was within the ORF region.

Actinomycetales↗
PubMed1995

Catalytic cleavage of an RNA target by 2-5A antisense and RNase L.

2-5A antisense (2-5A-AS) molecules are chimeric oligonucleotides that cause 2-5A-dependent RNase (RNase L) to catalyze the selective cleavage of RNA in human cells. These composite nucleic acids consist of a 5'-monophosphorylated, 2',5'-linked oligoadenylate known as 2-5A (an activator of RNase L) covalently attached to antisense 3',5'-oligodeoxyribonucleotides. Here, we characterize the targeted cleavage of the double-stranded RNA-dependent protein kinase (PKR) mRNA by purified, recombinant human RNase L. A 2-5A-AS chimera, which contains complementary sequence to PKR mRNA, and unmodified 2-5A, which causes general RNA decay, were about 20- and 40-fold more active, respectively, than 2-5A-AS chimeras in which the DNA domains are not complementary to sequences in PKR mRNA. Directed cleavage was efficient because each 2-5A-AS chimera targeted many RNA molecules. Moreover, RNase L caused the catalytic cleavage of the RNA target (kcat of approximately 7 s-1). The precise sites of PKR mRNA cleavage caused by 2-5A-AS were mapped, using a primer extension assay, to phosphodiester bonds adjacent to the 3' terminus of the chimera binding site (5' on the RNA target) as well as within the chimera's oligonucleotide binding site itself. The selectivity of this approach is shown to be provided by the antisense arm of the chimera, which places the RNA target in close proximity to the RNase.

Base Sequence↗
PubMed1995

Expression of the human MGMT O6-methylguanine DNA methyltransferase gene in a yeast alkylation-sensitive mutant: its effects on both exogenous and endogenous DNA alkylation damage.

Common Mer- cell lines deficient in O6-methylguanine DNA methyltransferase (MTase) activity probably result from the down-regulation of, rather than mutations in, the MGMT gene. However, the down-regulation of other unrelated genes was also observed in some of these cell lines, making it difficult to determine the precise functions of the MGMT MTase gene. To study the biological function of human MGMT MTase, we seek to utilize a newly created yeast mgt1 mutant deficient in the DNA repair MTase activity. The human MGMT cDNA was cloned into yeast expression vectors so that the MGMT gene is under the control of either an inducible GAL1 promoter or a constitutive ADH1 promoter. Upon galactose induction, the PGAL1-MGMT transformant had about 40-fold MTase activity compared to the wild-type strain. MGMT overexpression protected the yeast mgt1 mutant against alkylation-induced killing and mutation. Limited expression of the MGMT gene in the mgt1 mutant still provides significant alkylation resistance, albeit at a reduced level. The yeast mgt1 mutants increase spontaneous mutation rate, whereas constitutive expression of the MGMT gene lowered the spontaneous mutation rate in the mgt1 mutant to the wild-type level. We suggest that MGMT MTase may play the same role in human cells as the MGT1 MTase in yeast cells. Thus our results demonstrate that the human MGMT gene functionally complements the yeast MTase-deficient mutant in the protection against exogenous and endogenous DNA alkylation damage, which provides a useful tool for the study of in vivo mammalian MTase functions.

Alkylation↗
PubMed1995

DNA mismatch repair mutants do not increase N-methyl-N'-nitro-N-nitrosoguanidine tolerance in O6-methylguanine DNA methyltransferase-deficient yeast cells.

Treatment of cells with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) produces, among other lesions, mutagenic and carcinogenic lesions such as O6-methylguanine (O6MeG) and O4-methylthymine in DNA. An O6MeG DNA methyl-transferase (MTase) specifically and efficiently repairs such lesions. MTase-deficient bacterial, yeast and mammalian cells exhibit increased sensitivity not only to MNNG-induced mutagenesis, but also to MNNG-induced killing, suggesting that O6MeG-type lesions are also lethal to the cells. The lethal effect caused by O6MeG is not clear. Results from several recent experiments indicate that some MNNG-tolerant cell lines exhibit a loss of DNA mismatch binding/repair activity, suggesting that functional mismatch repair is probably responsible for the cellular sensitivity to DNA methylating agents. We tested this abortive O6MeG-T mismatch repair hypothesis in a well-defined lower eukaryote, Saccharomyces cerevisiae. We found that while mgt1-deleted MTase-deficient yeast strains are hypersensitive to MNNG-induced killing, combination of this mutation with any of the mlh1, msh2 or pms1 mutations did not render cells more tolerant to killing. msh3 mutation also did not rescue MNNG-induced genotoxicity. Furthermore, through the isolation and characterization of MNNG-tolerant cell lines from the MTase-deficient mutants we demonstrated that a DNA mismatch repair defect is neither sufficient nor required for this process. Since both DNA repair MTases and mismatch repair proteins are highly conserved between yeast and mammalian cells, our results could suggest alternative mechanisms in the cellular tolerance to O6MeG-induced killing.

Cells, Cultured↗
PubMed1995

Papilloma formation in human foreskin xenografts after inoculation of human papillomavirus type 16 DNA.

A mouse model of high-risk human papillomavirus infection was developed in which human papillomavirus (HPV) type 16 DNA was inoculated into human foreskin grafted to the skin of severe combined immunodeficient (scid) mice. Grafted skin contained human epidermis and dermis and, like normal human skin, expressed involucrin in differentiating keratinocytes. HPV type 16 DNA, attached to gold particles, was delivered directly into human epidermal cells and induced exophytic papilloma with histologic features of papillomavirus infection, including koilocytosis and expression of papillomavirus capsid antigen. This model should be useful for determining in vivo the functions of viral genes and for developing strategies to prevent and treat HPV-associated disease. It may also be of value in developing animal models of other human skin diseases.

Animals↗
PubMed1995

[Modern history of gynecology in China].

Dealing with the modern history (1850-1949) of the establishment and development of Gynecology in China, this paper claims that the sprouting period covers the time prior to 1899. After 1991 Gynecology developed steadily due to the advancement of surgery, pathology, bacteriology and anesthesiology. During this period, there were definite improvements in Gynecology diagnosis, treatment of pelvic inflammation and gynecological endocrinology.

China↗
PubMed1995

[History of modern obstetrics in western medicine in china].

This paper deals with the developmental history of western medicine in China. Due to the hindrance of feudal conception, modern obstetrics began rather late. The first Chinese obstetrical ward was established in Fujian Province in 1911. Before 1930, the Chinese maternal and neonatal mortality rates were 4-5 times higher than that of European and American countries. After 1930, "Education of Midwives" commenced, including the founding of midwifery school, antenatal care, pelvie measurements, improvements of the diagnosis for pathological pregnancy, management of complications during pregnancy. All these have improved modern obstetrics in China and laid down the foundation for its development after 1949.

China↗
PubMed1995

The factors affecting transformation efficiency of coryneform bacteria by electroporation.

High-voltage electroporation was performed to transfer plasmid DNA of pXZ10145 into different kinds of corynebacteria strains. A number of factors that affected transformation efficiency were investigated. Cells grown in the presence of 4% glycine and harvested in the early exponential growth phase (OD600 was about 0.2) were much more easily transformed. Transformation efficiency up to 8 x 10(6) transformants per microgram of plasmid DNA with homologously derived DNA was obtained. If heterologously derived DNA was used, transformation efficiency was 10(2) to 10(3) times lower than the former one.

Actinomycetales↗
PubMed1994

Transpogenes: the transposition-like integration of short sequence DNA into the yeast 2 micron plasmid creates the STB locus and plasmid-size polymorphism.

The type-2 2 mu plasmid of industrial yeast strains exhibits extensive size polymorphism in the STB (plasmid stability) locus and IR (inverted repeat)-right region. Comparative DNA sequence analyses of STB alleles identified a 38-bp sequence flanked by a 25-bp direct repeat as the underlying structural motif. Variable unequal recombination within the direct repeat accounted for the observed polymorphism of STB alleles. IR-right polymorphism was observed to result from tandem duplication of a 22-bp sequence flanked by a 9-bp direct repeat. The flanking direct repeats marked both loci as originating from the transposition-like integration of short DNA fragments. We call these structures transpogenes and note that these are hybrid structures of host and foreign DNA which can evolve into functional loci.

Base Sequence↗
PubMed1994

Blockage of NF-kappa B signaling by selective ablation of an mRNA target by 2-5A antisense chimeras.

Activation of 2-5A-dependent ribonuclease by 5'-phosphorylated, 2',5'-linked oligoadenylates, known as 2-5A, is one pathway of interferon action. Unaided uptake into HeLa cells of 2-5A linked to an antisense oligonucleotide resulted in the selective ablation of messenger RNA for the double-stranded RNA (dsRNA)-dependent protein kinase PKR. Similarly, purified, recombinant human 2-5A-dependent ribonuclease was induced to selectively cleave PKR messenger RNA. Cells depleted of PKR activity were unresponsive to activation of nuclear factor-kappa B (NF-kappa B) by the dsRNA poly(I):poly(C), which provides direct evidence that PKR is a transducer for the dsRNA signaling of NF-kappa B.

Adenine Nucleotides↗
PubMed1994

Femtosecond pump-probe analysis of energy and electron transfer in photosynthetic membranes of Rhodobacter capsulatus.

Low-intensity, 295 K, femtosecond pump-probe transient absorption measurements are described that have been performed to investigate energy and electron transfer in photosynthetic membranes from a Rhodobacter capsulatus strain lacking functional light harvesting antenna complex II. Spectral and kinetic similarities between the absorption changes of isolated reaction centers and those of reaction centers in membranes upon 800-nm excitation suggest that the charge separation process in both cases is very similar. An ultrafast energy relaxation process observed near 872 nm when 800-nm excitation is used is interpreted as interexcitonic relaxation within the antenna, though other interpretations, such as vibrational relaxation, are possible. On the basis of global exponential fitting analysis of the time-dependent spectral changes using 800- and 880-nm excitation wavelengths to selectively excite the reaction center and the LHI antenna, respectively, it is found that excitation energy transfer and trapping in Rb. capsulatus is limited by the overall rate of energy transfer between the antenna and the reaction center. This conclusion is supported by the observation that excitation at 800 nm, but not 880 nm, results in absorbance changes indicative of charge separation with a lifetime (3.1 ps) very close to that reported for charge separation in isolated reaction centers (3.5 ps). Thus, most reaction centers that are directly excited undergo charge separation and not backward energy transfer to the LHI antenna complexes. Both a kinetic model analysis and a direct comparison between time-resolved spectra obtained using different excitation wavelengths resulted in an energy-detrapping efficiency of about 15 +/- 10%.

Bacterial Proteins↗
PubMed1994

The MAG1* 3-methyladenine DNA glycosylase gene is closely linked to the SPT15 TATA-binding TFIID gene on chromosome V-R in Saccharomyces cerevisiae.

The MAG1 gene encodes a 3-methyladenine DNA glycoslyase, which is involved in DNA alkylation repair in Saccharomyces cerevisiae. The mag1 mutant is deficient in 3-methyladenine DNA glycosylase activity and shows enhanced sensitivity to several monofunctional alkylating agents. MAG1 is allelic to MMS5. This gene has been previously located on chromosome V by chromosomal hybridization. We present physical and genetic mapping data here showing that the MAG1 gene is located on chromosome V-R, proximal to and about 10 kilobase pairs away from the SPT15 gene coding for the yeast TATA-binding protein TFIID.

Chromosome Mapping↗