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Biomedical subjects

Y Cao

Publications and source records attributed to Y Cao.

At least 505 records · Page 28Linked to original sources

An infrequent point mutation of the p53 gene in human nasopharyngeal carcinoma.

Point mutations in the p53 gene have been detected in a variety of human cancers; the mutations are clustered in four "hot-spots" located in the coding region of exons 5, 7, and 8, which coincide with the four most highly conserved regions of the gene. We report the finding of a heterozygous G----C mutation at codon 280 (exon 8), position 2, of the p53 gene in a nasopharyngeal carcinoma (NPC) cell line, originating from Guangdong, a province in the People's Republic of China that leads the world in NPC incidence. A survey of nasopharyngeal tissues and NPC biopsies revealed that 1 out of 12 NPC samples from Hunan, another province in the People's Republic of China with high NPC incidence, had the same heterozygous mutation at codon 280 of p53, and none of 10 biopsies from Taiwan showed a mutation within exons 5-8 of the p53 gene. No other alteration of gene structure, including gross rearrangement or loss of heterozygosity or abnormality of gene expression was detected in NPC cell lines or NPC biopsies. We conclude from this study that mutational or other alterations of the p53 gene are not common in nasopharyngeal carcinogenesis and that a codon-280 mutation of p53 may be involved in less than 10% of NPC cases. This result contrasts with the relatively high frequency of p53 mutations associated with several other human carcinomas and suggests the importance of other genes in NPC genesis.

Base Sequence↗

Analysis of gene expression in single live neurons.

We present here a method for broadly characterizing single cells at the molecular level beyond the more common morphological and transmitter/receptor classifications. The RNA from defined single cells is amplified by microinjecting primer, nucleotides, and enzyme into acutely dissociated cells from a defined region of rat brain. Further processing yields amplified antisense RNA. A second round of amplification results in greater than 10(6)-fold amplification of the original starting material, which is adequate for analysis--e.g., use as a probe, making of cDNA libraries, etc. We demonstrate this method by constructing expression profiles of single live cells from rat hippocampus. This profiling suggests that cells that appear to be morphologically similar may show marked differences in patterns of expression. In addition, we characterize several mRNAs from a single cell, some of which were previously undescribed, perhaps due to "rarity" when averaged over many cell types. Electrophysiological analysis coupled with molecular biology within the same cell will facilitate a better understanding of how changes at the molecular level are manifested in functional properties. This approach should be applicable to a wide variety of studies, including development, mutant models, aging, and neurodegenerative disease.

Animals↗

Expression of an outward-rectifying potassium channel from maize mRNA and complementary RNA in Xenopus oocytes.

Injection of Xenopus oocytes with poly(A)+ mRNA isolated from different plants (maize, cucumber, and squash) results in the appearance of a voltage- and time-dependent, potassium-selective, outward current that is similar to the outward-rectifying potassium current recorded in many higher plant cells. Maize shoots were found to be especially enriched in mRNA encoding such activity. A cDNA library of maize shoot mRNA was constructed in the vector lambda ZAPII and was used to synthesize RNA complementary to the cDNA (cRNA). Injection of the cRNA gave rise to an outward-rectifying potassium current with properties similar to the currents obtained by poly(A)+ mRNA injection. These results demonstrate that higher plant mRNA can be properly translated into a product that produces a voltage-regulated potassium channel in the plasma membrane of Xenopus oocytes. Thus, Xenopus oocytes can be used as a heterologous expression system for the functional identification and isolation of plant ion channel genes as well as for the study of structure-function relationship of plant ion channels.

Animals↗

The two consecutive M substates in the photocycle of bacteriorhodopsin are affected specifically by the D85N and D96N residue replacements.

The photocycle of the proton pump bacteriorhodopsin contains two consecutive intermediates in which the retinal Schiff base is unprotonated; the reaction between these states, termed M1 and M2, was suggested to be the switch in the proton transport which reorients the Schiff base from D85 on the extracellular side to D96 on the cytoplasmic side (Váró and Lanyi, Biochemistry 30, 5016-5022, 1991). At pH 10 the absorption maxima of both M1 and M2 could be determined in the recombinant D96N protein. We find that M1 absorbs at 411 nm as do M1 and M2 in wild-type bacteriorhodopsin, but M2 absorbs at 404 nm. Thus, in M2 but not M1 the unprotonated Schiff base is affected by the D96N residue replacement. The connectivity of the Schiff base to D96 in the detected M2 state, but not in M1, is thereby established. On the other hand, the photostationary state which develops during illumination of D85N bacteriorhodopsin contains an M state corresponding to M1 with an absorption maximum shifted to 400 nm, suggesting that this species in turn is affected by D85. These results are consistent with the suggestion that M1 and M2 are pre-switch and post-switch states, respectively.

Bacteriorhodopsins↗

Laminin on Toxoplasma gondii mediates parasite binding to the beta 1 integrin receptor alpha 6 beta 1 on human foreskin fibroblasts and Chinese hamster ovary cells.

We investigated the role of parasite-bound laminin and the host cell beta 1 integrin receptors for this extracellular matrix protein in Toxoplasma gondii binding to fibroblasts. Laminin but not fibronectin was detected on extracellular tachyzoites by immunofluorescence and immunoblotting. Binding of parasites to CHO cells was inhibited by polyclonal antibodies to laminin and by a monoclonal antibody directed against the globular carboxyl-terminal portion of the long arm of laminin (at or near the suggested ligand-binding sites for alpha 3 beta 1 and alpha 6 beta 1), but not by a monoclonal antibody directed against the lateral short arms of laminin near the cross region of the molecule. Antibodies to the alpha 6 but not the alpha 2, alpha 3, or alpha 5 chains of the beta 1 family of integrins blocked parasite attachment to human foreskin fibroblasts and CHO cells. Attachment of T. gondii to cells via laminin on the parasite surface and laminin receptors on the mammalian cell is consistent with the capacity of the parasite to invade almost all nucleated cells.

Animals↗

Construction of a recombinant oral vaccine against Salmonella typhi and Salmonella typhimurium.

The viaB gene coding for the Vi antigen of Salmonella typhi Ty2 was subcloned into expression vector pYA248. The recombinant plasmid was termed SMM202 and transformed into Salmonella typhimurium chi 4072, an attenuated delta cya delta crp mutant. Recombinant S. typimurium Vi4072 had the ability to produce Vi capsular polysaccharide and also to invade and colonize the small intestine, mesenteric lymph nodes, and spleen of BALB/c mice. Mice orally immunized with Vi4072 developed serum and secretory antibody responses to the Vi antigen, as measured by a passive hemagglutination assay. Mice developed a delayed-type hypersensitivity following a footpad injection with Vi antigen after being sensitized orally with a suitable dose of Vi4072. Immunization of mice with Vi4072 afforded complete protection against fatal infection with virulent S. typhi Ty2. All data indicate that this route of antigen delivery is effective for stimulating antibody-mediated immunity and for inducing a cell-mediated immune response in BALB/c mice. Thus, S. typhimurium Vi4072 may serve as a vaccine for protection against typhoid fever and salmonellosis caused by S. typhimurium.

Administration, Oral↗

[Effects of moxibustion on experimental gastric ulcer in rats].

This study was designed to determine whether moxibustion at "Shenjue" plays a role in protecting gastric mucosa in rats. The areas and histological changes of gastric ulcer were examined. The results were as follows: 1. Moxibustion pretreatment within 3 weeks significantly prevented the formation of gastric ulcer. 2. As gastric ulcer had formed, moxibustion couldn't shorten the period of recovery of gastric ulcer. The results suggested that moxibustion pretreatment at "Shenjue" had a protective effect on gastric mucosa. After the experimental ulcer had formed, however, the moxibustion had little protective effect on it.

Acetates↗

Behavioral effects of stem-leaves extract from panax ginseng C.A. Meyer.

The behavioral effects of extracts from ginseng stem and leaves (GL), standardized with respect to the total saponines, and from ginseng roots (G115), standardized with respect to the content of ginsenosides were examined in experiments on rats with undisturbed memory and in rats with experimentally-impaired memory (electroconvulsive shock) using the methods for active avoidance (shuttle-box) and passive avoidance (step-down, step-through), the water-maze method and the method for studying exploratory behavior. On multiple administration G115 exerted favorable effects on learning and memory and on the higher nervous activity as a whole. These effects greatly varied with the dose and administration schedules, with the rat strain, with the rat's ability to perform adequately in any particular learning task, and with the behavioral method. The extract from the overground part of ginseng (GL) had, in the majority of cases, an effect weaker than that of G115 or was without effect at all. Based on previous and present results, we discuss the role of the changes in brain biogenic monoamines induced by the extracts for their mechanism of action.

Amnesia↗

[Study on radionuclide migrating along the channel].

The study of impedance line along channel had been performed with the radionuclide tracing method in 15 healthy human being and 4 guinea pigs. These images of tracer migrating routes approximately corresponded with classical courses of channel. The study proved that these line-shaped tracer migration were not by the way of lymphatic and blood vessels as well as nerves. This image of radionuclide migrating along channel may used as the guide for the observation of micro-autoradiography. In this study, satisfactory migrating image along the channel was also obtained by 131I instead of 99mTC. So a new way was opened up for both macro- and micro-autoradiographic study.

Adult↗

[Changes in cardiac contractility induced by stimulation of hypothalamic ventromedial nucleus].

Experiments were performed on anesthetized cats to observe the effect of electrical stimulation of the hypothalamic ventromedial nucleus (VMH) on cardiac contractility as judged by the change in LVSP, dp/dtmax, Vmax, P-dp/dt, P-lndp/dt. Electrical stimulation of VMH induced an augment response in cardiac force, while stimulation of an area between VMH and the third ventricle led only to a decrease in cardiac contractile force. All these changes were not accompanied by significant changes in heart rate and unaffected by bilateral vagotomy. The present experiment indicates that the inotropic and chronotropic regulation centers of cardiac function exist in different hypothalamic areas.

Animals↗

Water is required for proton transfer from aspartate-96 to the bacteriorhodopsin Schiff base.

During the M in equilibrium with N----BR reaction sequence in the bacteriorhodopsin photocycle, proton is exchanged between D96 and the Schiff base, and D96 is reprotonated from the cytoplasmic surface. We probed these and the other photocycle reactions with osmotically active solutes and perturbants and found that the M in equilibrium with N reaction is specifically inhibited by withdrawing water from the protein. The N----BR reaction in the wild-type protein and the direct reprotonation of the Schiff base from the cytoplasmic surface in the site-specific mutant D96N are much less affected. Thus, it appears that water is required inside the protein for reactions where a proton is separated from a buried electronegative group, but not for those where the rate-limiting step is the capture of a proton at the protein surface. In the wild type, the largest part of the barrier to Schiff base reprotonation is the enthalpy of separating the proton from D96, which amounts to about 40 kJ/mol. We suggest that in spite of this D96 confers an overall kinetic advantage because when this residue becomes anionic in the N state its electric field near the cytoplasmic surface lowers the free energy barrier of the capture of a proton in the next step. In the D96N protein, the barrier to the M----BR reaction is 20 kJ/mol higher than what would be expected from the rates of the M----N and N----BR partial reactions in the wild type, presumably because this mechanism is not available.

Aspartic Acid↗

Molecular abnormalities in the expression of HLA class-I antigens by melanoma cells.

Testing of a panel of cultured human melanoma cells with radiolabelled anti-HLA-class-I monoclonal antibodies (MAbs) in a binding assay has shown lack of reactivity of FO-I and SK-MEL-33 cells and low reactivity of SK-MEK-19 cells. SDS-PAGE analysis of the components immunoprecipitated from the 3 intrinsically radiolabelled melanoma cell lines by antibodies to the 2 subunits of HLA-class-I antigens has not detected beta 2-mu in the immunoprecipitates from melanoma cells FO-I and SK-MEL-33 and only a low level of HLA-class-I heavy chain in the immunoprecipitate from SK-MEL-19 cells. Northern blotting analysis with probes specific for HLA-class-I heavy chain and for beta 2-mu indicates that the abnormalities in HLA-class-I-antigen expression reflects a defect at the transcriptional level in FO-I cells and at the post-transcriptional level in SK-MEL-19 and in SK-MEL-33 cells. FO-I, SK-MEL-19 and SK-MEL-33 cells represent useful models to analyze the molecular mechanisms underlying the loss of HLA-class-I-antigen expression which is often associated with malignant transformation of melanocytes and to characterize the role of HLA-class-I antigens in the biology of melanoma cells and in their interactions with effector cells.

Antibodies↗

Isolation and partial characterization of a transformation-associated sequence from human nasopharyngeal carcinoma.

A transforming activity associated with Chinese nasopharyngeal carcinoma (NPC) cell line CNE2 DNA has been identified by transfer into nontransformed promotion-sensitive mouse JB6(P+) C141 cells. To clone this transformation-associated sequence, we carried out three cycles of transfection, followed by cloning of anchorage-independent transformants in soft agar. A tertiary CNE/JB6 clonal transfectant cell line 625 whose DNA showed transforming activity, as indicated in both soft-agar assay and nude-mice implantation, was used to make a genomic library in the vector lambda dash. Using the human repeated sequence Blur 8 to screen the library, we obtained 10 human Alu-positive clones. A cloned Alu-positive insert of 16 kbp, CNE 323, was characterized in detail. CNE 323 transferred moderate transforming activity when introduced into JB6 P+ cells and showed no homology to Ha-, Ki-, or N-ras genes; human promotion sensitivity genes; src, myb, jun, myc, fos, raf, or int-2 oncogenes; or epidermal growth factor receptor. The isolated CNE 323 DNA sequence appeared to preserve the genomic structure of the original sequence found in CNE2 cells and in nude mouse tumors induced by CNE2 cells or by CNE/JB6 transfectant cells, indicating that the cloned NPC sequence was activated during NPC carcinogenesis and not during transfection or construction of the library, and that the cloned sequence or a larger sequence of which it was part played a role in tumor formation. Finally, we identified a 1.3-kb mRNA that hybridizes to a subclone of the 16-kb NPC sequence in CNE2 cell poly (A)+ RNA.

Animals↗

Glycogen metabolism and signal transduction in mammals and yeast.

Mammalian glycogen synthase, with its complex multisite phosphorylation mechanisms, continues to provide interesting and novel examples of the regulation of protein function. The mammalian enzyme is phosphorylated in a hierarchal manner such that modification of certain sites requires the prior phosphorylation of other sites. Yeast contains two glycogen synthases that have extensive similarities to their mammalian counterpart but the greatest divergence in amino acid sequence is seen precisely in the regions likely to be involved in covalent control. We hope that examination of the control of the yeast glycogen synthase will be as informative as study of the mammalian enzymes, whether by revealing important parallels with the mammalian system or by uncovering major differences in mechanism.

Amino Acid Sequence↗