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Biomedical subjects

Jia Liu

Publications and source records attributed to Jia Liu.

At least 73 records · Page 4Linked to original sources

Regulation of a novel cell differentiation-associated gene, JWA during oxidative damage in K562 and MCF-7 cells.

Oxidative stress, or the production of oxygen-centered free radicals, has been hypothesized as the major source of DNA damage that can lead to a variety of diseases including cancer. It is known that 8-hydroxy-deoxyguanosine (8-oxo-dG) is a useful biomarker of oxidative DNA damage. Our recent data showed that JWA, initially being cloned as a novel cell differentiation-associated gene, was also actively responsive to environmental stressors, such as heat-shock, oxidative stress and so on. In the present study, we have applied a modified comet assay and bacterial repair endonucleases system (endonuclease III and formamidopyrimidine glycosylase) to investigate if JWA is involved in hydrogen peroxide (H2O2)-induced DNA damage and repair in K562 and MCF-7 cells, and to demonstrate if the damage is associated with 8-oxo-dG. The results from the comet assay have shown that the average tail length and the percentage of the cells with DNA tails are greatly induced by H2O2 treatment and further significantly enhanced by the post-treatment of repair endonucleases. The H2O2-induced 8-oxo-dG formation in K562 and MCF-7 cells is dose-dependent. In addition, the data have clearly demonstrated that JWA gene expression is actively induced by H2O2 treatment in K562 and MCF-7 cells. The results suggest that JWA can be regulated by oxidative stress and is actively involved in the signal pathways of oxidative stress in the cells.

8-Hydroxy-2'-Deoxyguanosine↗

Stereospecific effects determine the structure of a four-way DNA junction.

Conversion of a centrally located phosphate group to an electrically neutral methyl phosphonate in a four-way DNA junction can exert a major influence on its conformation. However, the effect is strongly dependent on stereochemistry. Substitution of the proR oxygen atom by methyl leads to conformational transition to the stacking conformer that places this phosphate at the point of strand exchange. By contrast, corresponding modification of the proS oxygen destabilizes this conformation of the junction. Single-molecule analysis shows that both molecules are in a dynamic equilibrium between alternative stacking conformers, but the configuration of the methyl phosphonate determines the bias of the conformational equilibrium. It is likely that the stereochemical environment of the methyl group affects the interaction with metal ions in the center of the junction.

DNA↗

The R1 resistance gene cluster contains three groups of independently evolving, type I R1 homologues and shows substantial structural variation among haplotypes of Solanum demissum.

Cultivated and wild potatoes contain a major disease-resistance cluster on the short arm of chromosome V, including the R1 resistance (R) gene against potato late blight. To explore the functional and evolutionary significance of clustering in the generation of novel disease-resistance genes, we constructed three approximately 1 Mb physical maps in the R1 gene region, one for each of the three genomes (haplotypes) of allohexaploid Solanum demissum, the wild potato progenitor of the R1 locus. Totals of 691, 919 and 559 kb were sequenced for each haplotype, and three distinct resistance-gene families were identified, one homologous to the potato R1 gene and two others homologous to either the Prf or the Bs4 R-gene of tomato. The regions with R1 homologues are highly divergent among the three haplotypes, in contrast to the conserved flanking non-resistance gene regions. The R1 locus shows dramatic variation in overall length and R1 homologue number among the three haplotypes. Sequence comparisons of the R1 homologues show that they form three distinct clades in a distance tree. Frequent sequence exchanges were detected among R1 homologues within each clade, but not among those in different clades. These frequent sequence exchanges homogenized the intron sequences of homologues within each clade, but did not homogenize the coding sequences. Our results suggest that the R1 homologues represent three independent groups of fast-evolving type I resistance genes, characterized by chimeric structures resulting from frequent sequence exchanges among group members. Such genes were first identified among clustered RGC2 genes in lettuce, where they were distinguished from slow-evolving type II R-genes. Our findings at the R1 locus in S. demissum may indicate that a common or similar mechanism underlies the previously reported differentiation of type I and type II R-genes and the differentiation of type I R-genes into distinct groups, identified here.

Chromosomes, Artificial, Bacterial↗

Analyzing the potato abiotic stress transcriptome using expressed sequence tags.

To further increase our understanding of responses in potato to abiotic stress and the potato transcriptome in general, we generated 20 756 expressed sequence tags (ESTs) from a cDNA library constructed by pooling mRNA from heat-, cold-, salt-, and drought-stressed potato leaves and roots. These ESTs were clustered and assembled into a collection of 5240 unique sequences with 3344 contigs and 1896 singleton ESTs. Assignment of gene ontology terms (GOSlim/Plant) to the sequences revealed that 8101 assignments could be made with a total of 3863 molecular function assignments. Alignment to a set of 78 825 ESTs from other potato cDNA libraries derived from root, leaf, stolon, tuber, germinating eye, and callus tissues revealed 1476 sequences unique to abiotic stressed potato leaf and root tissue. Sequences present within the 5240 sequence set had similarity to genes known to be involved in abiotic stress responses in other plant species such as transcription factors, stress response genes, and signal transduction processes. In addition, we identified a number of genes unique to the abiotic stress library with unknown function, providing new candidate genes for investigation of abiotic stress responses in potato.

Arabidopsis↗

[Down-regulation of caveolin-1 in gastric carcinoma and its clinical biological significance].

BACKGROUND & OBJECTIVE: Caveolin-1, a candidate tumor suppressor gene, aberrantly expressed in many kinds of carcinomas. This research was designed to check the expression of Caveolin-1 in non-cancerous gastric mucosa, intestinal metaplasia, atypical hyperplasia, gastric cancer tissues, and gastric cancer cell lines MGC803 and BGC823, and to analyze its clinical biological significance in stepwise gastric carcinogenesis. METHODS: Frozen gastric tissue array-based immunohistochemistry (IHC) was used to examine the expression of Caveolin-1 in 56 specimens of gastric cancer, 29 specimens of non-cancerous mucosa, 11 specimens of intestinal metaplasia, and 7 specimens of atypical hyperplasia. Correlations of Caveolin-1 expression to clinical stage, lymph node metastasis, Lauren's type, and histological type were analyzed. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect mRNA and protein levels of Caveolin-1 in 56 specimens of gastric cancer and relevant adjuvant non-cancerous tissue, and cell lines MGC803 and BGC823. RESULTS: Positive rate of Caveolin-1 was significantly lower in gastric cancer than in non-cancerous mucosa, intestinal metaplasia, and atypical hyperplasia (17.9% vs. 84.8%, 81.8%, and 57.1%, P<0.05). Positive rate of Caveolin-1 was lower in advanced gastric cancers than in gastric cancer of early stage (16.0% vs. 33.3%), but the difference was not significant (P>0.05). Positive rate of Caveolin-1 was significantly lower in diffuse gastric cancer than in gastric cancer of intestinal type (7.0% vs. 26.9%, P<0.05), significantly lower in the cases with lymph node metastases than in the cases without lymph node metastases (9.7% vs. 31.8%, P<0.05). Protein and mRNA levels of Caveolin-1 in gastric cancer and relevant adjuvant non-cancerous tissues have no significant difference, but its expression was low in MGC803 and BGC823 cells. CONCLUSION: Progressive down-regulation of Caveolin-1 in gastric epithelial cells is correlated to gastric carcinogenesis.

Caveolin 1↗

Regulatory effects of emodin on NF-kappaB activation and inflammatory cytokine expression in RAW 264.7 macrophages.

Emodin, an anthraquinones component of Rheum palmatun, has been used for anti-inflammatory purposes. However, its underlying molecular effect(s) on target cells remain to be well clarified. Thus, our current study was aimed at investigating the regulatory mechanism of emodin on liposaccharide-induced inflammatory responses in RAW 264.7 macrophages by RT-PCR, Western blot analysis, immunocytochemical staining and immunofluorescence analysis. It was found that a treatment of 20 microg/ml emodin inhibited the expression of a panel of inflammatory-associated genes, including TNFalpha, iNOS, IL-10, cytosolic IkappaBalpha, IKK-alpha and IKK-gamma, to different extents as well as the nuclear translocation of NF-kappaB (nuclear factor-kappaB). The promoting effect of emodin on the production and translocation of p105 (the precursor of NF-kappaB p50) was time-dependent and reached a maximum at 5 h. Our data suggest that emodin plays its anti-inflammatory roles by regulating inflammatory cytokines, specifically by suppressing NF-kappaB activation.

Active Transport, Cell Nucleus↗

[Zona pellucida and male fertilization].

Zona pellucida (ZP) plays a key role in the recognition, combination and penetration of sperms and oocytes, the prevention of multiple impregnation, the protection of embryos, and so on. The paper reviews the constitution, molecular structure, biological function and application of zona pellucida in male fertilization.

Acrosome Reaction↗

[The role of reactive oxygen species in N-[4-hydroxyphenyl] retinamide induced apoptosis in bladder cancer cell lineT24].

OBJECTIVE: To study the mechanism of the apoptosis induced by N-[4-hydroxyphenyl] retinamide (4-HPR) in bladder cancer cell line T24, and the involvement of DNA damage and repair. METHODS: T24 cells were treated with 4-HPR at the concentration of 2.5, 5.0 and 10.0 micromol/L, and the cell grow inhibition was measured by cell counting assay. The fluorescent intensity of reactive oxygen species (ROS) was determined by spectrofluorometer. The apoptosis was measured by flow cytometry and DNA fragment assay. The expression of XRCC1 protein and activation of caspase-3 were detected by Western blot. RESULTS: 4-HPR induced apoptosis in T24 cell. A dose-dependent increase in the percentage of apoptosis cells was observed (1.8%, 4.0% and 10.5% respectively at 2.5, 5.0, 10.0 micromol/L 4HPR). In the meantime, ROS level in the cell was increased (peaked at 3 fold). It also caused down-regulation of the expression of XRCC1, and activation of caspase-3. Vitamin C effectively inhibited ROS rise induced by 4-HPR, and also partially inhibited cell growth, apoptosis, and down-regulation of the expression of XRCC1. CONCLUSION: The generation of ROS and DNA damage may be the major mechanism of the apoptosis of bladder cancer cell line T24 induced by 4-HPR.

Antineoplastic Agents↗

[Study on percutaneous absorption of Xuetie Dingchuan Plaster].

OBJECTIVE: To filtrate prescription of the adhesive plaster, kind and amount of percutaneous absorption reagent, etc. METHODS: In vitro percutaneous absorption of Xuetie Dingchuan Plaster was studied by HPLC using magnolin as the content of the marker substance, in the condition of equal effective component of samples, drawing graphs with time as abscissa and area of peak as ordinate. RESULTS: 40% ethanol was used as receptor liquid, prescription with Polyethylene Glycol 6000 as bases, azone as percutaneous absorption reagent. The amount of azone was 1.8%. CONCLUSION: The prescription of Xuetie Dingchuan Plaster, with 1.8% azone as percutaneous absorption reagent, and Polyethylene Glycol 6000 as bases, is preferable.

Administration, Cutaneous↗

[Study on the relationship between polymorphisms of Cyp1A1, GSTM1, GSTT1 genes and the susceptibility to acute leukemia in the general population of Hunan province].

OBJECTIVE: Based on the distribution of genetic polymorphisms regarding phase I metabolic enzyme cytochrome P450 1A1 (CYP1A1) and phase II metabolic enzymes glutathione S-transferase GSTM1 and GSTT1 genotypes in acute leukemia patients and health controls among general population of Hunan in China, this study was to explore the relationship between these gene polymorphisms and the susceptibility to acute leukemia. METHODS: Using case-control methodology, we studied 204 healthy controls and 232 patients with acute leukemia, of which 112 patients were suffering acute lymphoblastic leukemia (ALL) and 120 with acute non-lymphoblastic leukemia (ANLL). The frequencies of the genotypes were detected by PCR and PCR-RFLP techniques. RESULTS: The variation frequencies of CYP1A1 gene (Msp I polymorphisms, site 3801T-C variation) in ALL and ANLL groups were 74.1% and 70.8% respectively which were higher than 63.3% appeared in the healthy controls. However, the differences between patients (ALL or ANLL) and healthy controls were not statistically significant (P > 0.05 for both). The null genotype of GSTM1 (GSTM1 -/-) in ALL group was 60.7%, which was not significantly different from the controls (55.4%). However, GSTM1 -/- genotype in ANLL group was 68.3%, significantly different from the controls (P < 0.05). The null genotypes among GSTT1 (GSTT1 -/-) in ALL, ANLL and control group were 50.9%, 55.0% and 49.0% but their differences were not statistically significant (P > 0.05). The incidences of GSTM1 -/- and GSTT1 -/- combined genotype in ALL, ANLL and control group were 33.0%, 40.0% and 27.5%, of which the difference between ANLL group and control group was statistically significant (P < 0.05) and CYP1A1 gene heterozygous mutation type or homozygous mutation type combined with GSTM1 -/- and GSTT1 -/- increased the risk of ANLL (OR value 1.890, 95% CI: 1.084-3.295). CONCLUSION: These results indicated that both the variation of CYP1A1 gene or GSTT1 -/- genotype alone might not be associated with the susceptibility of acute leukemia while GSTM1 -/- genotype alone or combined with GSTT1 -/- or the 3801 T-C variation of CYP1A1 gene were correlated with ANLL. These findings suggest that GSTM1 - / - genotype alone or in combination with other defective genotypes might serve as risk factors to the etiology of ANLL.

Case-Control Studies↗

Correlation of Wnt-2 expression and beta-catenin intracellular accumulation in Chinese gastric cancers: relevance with tumour dissemination.

Wnt/beta-catenin signalling pathway is integrally associated with human tumour development and progression. Aberrant beta-catenin intracellular distribution has been found in gastric cancer, but the pattern of Wnt expression in stepwise gastrocarcinogenesis and its potential influence in beta-catenin distribution are still lesser known. By the methods of frozen tissue array-based immunohistochemistry, Western blot analysis and RT-PCR, a paralleled study was conducted to check Wnt2 expression and beta-catenin intracellular distribution in two major subtypes of gastric cancers (intestinal gastric cancer, i-GC and diffuse gastric cancer, d-GC) and their premalignant (intestinal metaplasia, IM and chronic gastritis, CG) and noncancerous counterparts. According to the results obtained and the clinical data collected, correlation of Wnt2 expression with beta-catenin translocalisation and their links with tumour dissemination were elucidated. The results demonstrated (1) that Wnt2 expression and cytoplasmic/nuclear beta-catenin accumulations appeared in most gastric cancers irrespective to their morphological phenotypes, (2) that over-expressed Wnt and nuclear translocalisation of beta-catenin were found in 68 and 58% of i-GCs and in 47 and 47% of d-GCs in a closely related pattern (P<0.01) and (3) that co-existence of Wnt2 up-regulation/beta-catenin nuclear translocalisation were positively associated with lymph node metastasis (P<0.05) as well as T-stage. These data indicate that Wnt/beta-catenin signalling pathway is activated in most of gastric cancers, which may play pivotal roles either in gastric cancer formation or in tumour invasion and dissemination.

Blotting, Western↗

Electrostatic interactions and the folding of the four-way DNA junction: analysis by selective methyl phosphonate substitution.

The structure and dynamics of the four-way (Holliday) junction are strongly dependent on the presence of metal ions. In this study, the importance of phosphate charge in and around the point of strand exchange has been explored by selective replacement with electrically neutral methyl phosphonate groups, guided by crystal structures of the junction in the folded, stacked X conformation. Junction conformation has been analysed by comparative gel electrophoresis and fluorescence resonance energy transfer (FRET). Three of sets of phosphate groups on the exchanging strands have been analysed; those at the point of strand exchange and those to their 3' and 5' sides. The exchanging and 3' phosphate groups form a box of negatively charged groups on the minor groove face of the junction, while the 5' phosphate groups face each other on the major groove side, with their proR oxygen atoms directed at one another. The largest effects are observed on substitution of the exchanging phosphate groups; replacement of both groups leads to the loss of the requirement for addition of metal ions to allow junction folding. When the equivalent phosphate groups on the continuous strands were substituted, a proportion of the junction folded into the alternative conformer so as to bring these phosphate groups onto the exchanging strands. These species did not interconvert, and thus this is likely to result from the alternative diasteromeric forms of the methyl phosphonate group. This shows that some of the conformational effects result from more than purely electrostatic interactions. Smaller but significant effects were observed on substitution of the flanking phosphate groups. All methyl phosphonate substitutions at these positions allowed folding to proceed at a reduced concentration of magnesium ions, with double substitutions more effective than single substitutions. Substitution of 5' phosphates resulted in a greater degree of folding at a given ionic concentration compared to the corresponding 3' phosphate substitutions. These results show that the phosphate groups at the point of strand exchange exert the largest electrostatic effect on junction folding, but a number of phosphate groups in the vicinity of the exchange region contribute to the overall effects.

DNA, Cruciform↗

Tetronamides as latent acyclic vinylogous amides in formal aza-[3 + 3] cycloaddition reactions with alpha,beta-unsaturated iminium salts. An unexpected rearrangement and an approach to synthesis of substituted piperidines.

A detailed account regarding formal aza-[3 + 3] cycloaddition reactions of tetronamides with alpha,beta-unsaturated iminium salts is described here. This investigation uncovers regioisomeric cycloadducts that were not found in previous studies involving this formal cycloaddition and an unexpected rearrangement that led to pyridines and dihydropyridines. Both stereochemical and regiochemical issues raised in this study provide further mechanistic insights into this cycloaddition. With careful control of reaction temperatures, the desired formal cycloadducts are obtained. Ensuing transformation of these cycloadducts into functionalized piperidines establishes the concept of employing tetronamides as latent acyclic vinylogous amides for the formal aza-[3 + 3] cycloaddition.

Amides↗

The M170 is selective for faces, not for expertise.

Are the mechanisms for face perception selectively involved in processing faces per se, or do they also participate in the processing of any class of visual stimuli that share the same basic configuration and for which the observer has gained substantial visual expertise? Here we tested the effects of visual expertise on the face-selective "M170", a magnetoencephalography (MEG) response component that occurs 170 ms after stimulus onset and is involved in the identification of individual faces. In Experiment 1, cars did not elicit a higher M170 response (relative to control objects) in car experts compared to controls subjects. In Experiment 2, the M170 amplitude was correlated with successful face identification, but not with successful car identification in car experts. These results indicate that the early face processing mechanisms marked by the M170 are involved in the identification of faces in particular, not in the identification of any objects of expertise.

Adult↗

Differential regulation of CYP1A1 and CYP1B1 expression in resveratrol-treated human medulloblastoma cells.

Resveratrol induces differentiation and Fas-independent apoptosis of medulloblastoma cells by a largely unknown mechanism. CYP1A1 and 1B1 are involved in resveratrol-mediated tumor suppression but their expression in medulloblastoma cells and their relevance to anti-medulloblastoma activity of resveratrol have not been described. The statuses of CYP1A1 and 1B1 in UW228-3 medulloblastoma cells without and with resveratrol treatments were elucidated in this study with ethoxyresorufin O-deethylation assay, followed by RT-PCR, immunocytochemical staining and Western blot hybridization. CYP1A1/1B1 enzymatic activity was low in UW228-3 cells but became several folds higher upon resveratrol treatments. CYP1A1 was undetectable and CYP1B1 was expressed in normally cultured cells. Accompanied by the increased fraction of apoptosis, enhanced CYP1A1 and downregulated CYP1B1 were observed in resveratrol-treated cells in time- and dose-related fashions. Our results demonstrate for the first time that in the medulloblastoma cell system, CYP1A1 upregulation is paralleled with resveratrol-induced differentiation and apoptosis, while CYP1B1 may not be an essential element in metabolic activation of resveratrol in those cells. CYP1A1 and 1B1 are resveratrol response genes and potential chemosensitive markers of medulloblastoma cells.

Antineoplastic Agents, Phytogenic↗

Effects of surface treatment and joint shape on microtensile bond strength of reattached root dentin segments.

STATEMENT OF PROBLEM: An effective bonding technique for reattaching vertical root fracture segments has not yet been established. PURPOSE: The purpose of this study was to investigate the effects of surface treatment and joint shape on bond strength of reattached root dentin segments. MATERIAL AND METHODS: Microtensile dentin specimens (n=120, cross-sectional area=1.5 mm2) were obtained from 12 bovine teeth roots, and fractured under a tensile force. The segments of each group (n=6) were reattached by a 4-META/MMA-TBB adhesive resin after different treatments (etching or polishing, a 50-microm or 500-microm gap space, butt or wedge joint). The group with a 50-microm gap space and butt joint, which was not etched or polished, served as the control. After 24 hours of storage in 0.9% normal saline solution or thermal cycling (3000 cycles, 4 degrees C and 60 degrees C), the specimens were fractured in a universal testing machine at a crosshead speed of 1.0 mm/min. The microtensile bond strengths (MPa) were recorded. The bonded interface and fractured surface were observed by SEM (original magnification x 1000). The results were analyzed by a 2-way analysis of variance and the Fisher protected least significant difference post-hoc test (alpha=.05). RESULTS: Significantly higher bond strengths (P=.0001) were shown for the nonpolished group (9.78 +/- 4.44 MPa) and the etched group (9.31 +/- 1.23 MPa) than for the polished group (fractured before test). The wedge-shaped joint exhibited the highest bond strength (18.57 +/- 3.40 MPa, P=.0017). SEM observations showed that the penetration of resin tags along the oblique dentinal tubules was within the wedge joint. CONCLUSION: Etching and oblique tubule orientation contributed to significantly high bond strength of reattached segments to dentin after vertical root fracture.

Acid Etching, Dental↗

Rhythm generation for food-ingestive movements.

In vitro block preparations of the central nervous system (CNS) are particularly valuable for study of central neuronal mechanisms controlling the respiratory and locomotor rhythms. No comparable in vitro preparation has been described previously, however, for analysis of analogous feeding rhythms. In this chapter, we present such a model. It is comprised of an in vitro brainstem-spinal cord preparation isolated from the newborn rat and mouse. Bath application to this preparation of N-methyl-D-aspartate (NMDA) induces rhythmical burst activity in the V, VII and XII nerves, which, collectively, is indicative of feeding behavior. Selected transections of the brainstem reveal that the central sucking rhythm generators for such V, VII and XII activity are separate from one another, and located segmentally in the brainstem at the level of their respective motor nuclei. We believe that use of this in vitro preparation will advance understanding of the central neuronal mechanisms controlling sucking and mastication, and the developmental transition from sucking to mastication.

Animals↗