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Biomedical subjects

Q Guo

Publications and source records attributed to Q Guo.

At least 37 records · Page 2Linked to original sources

[Effect of free fatty acids on glucose transporter 4 and insulin signaling proteins in rat skeletal muscle cells].

OBJECTIVE: To investigate the effect of free fatty acids on glucose transpoter4 (GLUT4) and insulin signaling proteins Grb2 and ERK2 in rat skeletal cells. METHODS: After the skeletal muscle cells of Sprague-Dawley rats were incubated with palmitate (0.25 mmol/L) or oleate (0.125 mmol/L) for 12, 24 and 36 hours, Western blotting was used to assess the levels of GLUT4 and insulin signaling proteins Grb2 and ERK2. The level of GLUT4 RNA was determined by dot blotting. RESULTS: The protein and RNA contents of GLUT4 were significantly decreased (P < 0.05) and the protein levels of Grb2 and ERK2 were decreased also compared with control in rat skeletal muscle cells after incubated with free fatty acids for 12, 24 and 36 hours (P < 0.05). CONCLUSION: Free fatty acids inhibit glucose metabolism and insulin activity in rat skeletal muscle cells by downregulating the expression of GLUT4 gene and protein and downregulating the protein expression of Grb2 and ERK2.

Adaptor Proteins, Signal Transducing↗

Par-4 is a synaptic protein that regulates neurite outgrowth by altering calcium homeostasis and transcription factor AP-1 activation.

Although Par-4 (prostate apoptosis response-4) is involved in initiation of neurodegenerative cascades associated with certain neurodegenerative disorders, normal physiological roles of Par-4 in neurons have remained elusive. It was recently reported that Par-4 protein levels could be regulated at translational level in synaptic terminals following apoptotic insults, suggesting that Par-4 might play a role in synaptic function. We report that Par-4 is a synaptic protein preferably localized in postsynaptic density (PSD). The expression of Par-4 in synaptosome preparations and PSDs are developmentally and regionally regulated. Synaptic Par-4 is enriched in the cerebral cortex and the hippocampus, but not in the cerebellum. In vitro as well as in vivo experiments demonstrate that the levels of synaptic Par-4 increase as the neurons mature. Overexpression of Par-4 in transfected PC12 cells inhibits nerve growth factor (NGF)-induced cellular differentiation and neurite outgrowth by a mechanism involving aberrant elevation of intracellular calcium levels and suppression of activation of the transcription factor AP-1. The actions of Par-4 were consistently blocked by co-expression of the dominant negative regulator of Par-4 activity (the leucine zipper domain of Par-4). Since the leucine zipper domain of Par-4 (Leu.zip) may mediate protein--protein interactions, the results indicate that the actions of Par-4 require its interaction with other protein(s) or dimerization with itself. These results suggest that Par-4 may play an important role in postsynaptic signal transduction and regulation of cellular pathways associated with cellular differentiation and neurite outgrowth. Identification of Par-4 as a novel synaptic protein may have significant implications in understanding the mechanisms of synaptic functions in physiological and pathological settings.

Animals↗

Ribozyme-mediated inhibition of caspase-3 activity reduces apoptosis induced by 6-hydroxydopamine in PC12 cells.

6-Hydroxydopamine (6-OHDA) is a neurotoxin used in the induction of experimental Parkinson's disease in both animals and PC12 cells, which are derived from rat pheochromocytoma tumors and have many properties similar to dopamine neurons. Biochemical and molecular approaches have shown that low doses of 6-OHDA induce apoptosis in PC12 cells and, in the processing of apoptosis, caspases are crucial mediators, and caspase inhibition is sufficient to rescue PC12 cells from apoptosis induced by 6-OHDA. However, because this caspase inhibition targets multiple caspases, it is not known whether a single caspase is primarily responsible for effecting cell death in this model. To assess the particular member (caspase-3) of the ced-3 family relevant to cell death and to position their activation within the apoptotic pathway, we constructed a hammerhead ribozyme directed against rat caspase-3, which could downregulate the expression of caspase-3 in vitro and in vivo, and transfer to PC12 cells. The results show that the ribozymes against caspase-3 could protect PC12 cells from apoptosis induced by low doses of 6-OHDA. The PC12 cell transfected with the ribozymes shows a significant decrease in caspase-3 activity compared with control cells at various time points. Parallel to the reduced caspase-3 protease activity, similar decreased levels of apoptotic cells and DNA fragmentation were also assessed by staining with Hoechst 33258 and ELISA, respectively. Overexpression of p35, a general caspase inhibitor, also protected PC12 cells from apoptosis. These results confirm that caspases play an important role in 6-OHDA-induced PC12 cell apoptosis and indicate that caspase-3 itself is one of the crucial mediators of neurotoxin-induced PC12 cell apoptosis.

Adrenergic Agents↗

Inhibitory effect of alpha-lipoic acid and its positively charged amide analogue on nitric oxide production in RAW 264.7 macrophages.

The aim of this study was to investigate the effect of the mitochondrial cofactor alpha-lipoic acid [R (+) LA] or its lipoamide analogue, 2-(N,N-dimethylamine) ethylamido lipoate [R (+) LA-plus], on nitric oxide (NO) production in RAW 264.7 macrophages. NO production from RAW 264.7 cells stimulated with 10 microg/mL of lipopolysaccharide and 50 U/mL of interferon-gamma was measured directly by electron spin resonance using spin-trapping techniques. R (+) LA or R (+) LA-plus was found to inhibit NO production at pharmacologically relevant concentrations. However, in a cell-free chemical system, neither R (+) LA nor R (+) LA-plus was able to directly scavenge NO. Furthermore, in the presence of 2.5 or 25 mM glucose, the inhibitory effects of R (+) LA and R (+) LA-plus on NO production were decreased markedly, while they showed more potent inhibitory effects in the presence of 2 microM rotenone or 5 microg/mL of antimycin A, inhibitors of mitochondrial complex I and complex III, respectively. Glucose, rotenone, or antimycin A alone resulted in an increase of NO production. These results suggest that NO production in macrophages can be regulated by glucose and mitochondrial respiration, and that modulation of NO production by lipoic acid or lipoamide analogues in inflammatory situations is attributed not to their radical scavenging activity but to their redox properties.

Amides↗

Prostate apoptosis response-4 enhances secretion of amyloid beta peptide 1-42 in human neuroblastoma IMR-32 cells by a caspase-dependent pathway.

Prostate apoptosis response-4 (Par-4) is a leucine zipper protein that promotes neuronal cell death in Alzheimer's disease (AD). Neuronal degeneration in AD may result from extracellular accumulation of amyloid beta peptide (Abeta) 1-42. To examine the effect of Par-4 on Abeta secretion and to reconcile amyloid/apoptosis hypotheses of AD, we generated IMR-32 cell lines that overexpress Par-4 and/or its leucine zipper domain. Overexpression of Par-4 did not significantly affect levels of the endogenously expressed beta amyloid precursor protein but drastically increased the Abeta(1-42)/Abeta(total) ratio in the conditioned media about 6-8 h after trophic factor withdrawal. Time course analysis of caspase activation reveals that Par-4 overexpression exacerbated caspase activation, which is detectable within 2 h after trophic factor withdrawal. Furthermore, inhibition of caspase activity by the broad spectrum caspase inhibitor BD-fmk significantly attenuated the Par-4-induced increase in Abeta 1-42 production. In addition, the effects of Par-4 on secretion of Abeta 1-42 were consistently blocked by co-expression of the leucine zipper domain, indicating that the effect of Par-4 on Abeta secretion may require its interaction with other protein(s). These results suggest that Par-4 increases secretion of Abeta 1-42 largely through a caspase-dependent pathway after apoptotic cascades are initiated.

Amyloid beta-Peptides↗

Hazard of smoking initiation by age among adolescents in Wuhan, China.

BACKGROUND: Knowledge about age of smoking initiation among adolescents in China is helpful for exploring cultural differences in adolescent smoking behavior and informative for global tobacco control. However, little has been documented on this issue. METHOD: Adolescents (6,473) attending grades 7, 8, and 9 completed the baseline survey of a longitudinal, randomized smoking prevention trial. Data were collected in classrooms with a paper-and-pencil questionnaire. A survival model was used in the statistical analysis. RESULTS: The hazard of smoking initiation for boys showed a pattern previously observed in the United States: very low (<2%) before 7 years of age, increasing rapidly after age 10, and peaking at 14-15 years of age. The hazard for girls was below or around 1% until 12 years of age before it increased. The hazard levels were similar for adolescents both in urban and in rural areas, but higher for those in grade 7 than in grades 8 and 9. CONCLUSIONS: Chinese boys in Wuhan, China, experienced a hazard pattern of smoking initiation by age similar to those observed in the United States, while Chinese girls there experienced a rather low risk of smoking initiation. The hazard pattern suggests that the best time for smoking prevention is between 10 and 15 years of age. Adolescents in lower grades are at higher risk of early smoking initiation, suggesting a potential cohort effect in adolescent smoking initiation in Wuhan, China.

Adolescent↗

Protein binding of procyanidins: studies using polyacrylamide gel electrophoresis and French maritime pine bark extract.

The application of PAGE to determine the interaction between procyanidins and proteins, as presented here, enables one to directly determine the binding of either a pure of a complex mixture of flavonoids to a particular protein. If the protein of interest is an enzyme, the combination of PAGE with quantitative activity measurements allows identifying whether a change in the enzyme activity is related to the binding. Data presented suggest that PBE and EGb 761 have protein-binding properties, which, in addition to their redox-based effects, could provide a biochemical basis for their action in biological systems.

Electrophoresis, Polyacrylamide Gel↗

Assessing molecular similarity/diversity of chemical structures by FT-IR spectroscopy.

FT-IR spectra have been investigated for their ability to distinguish compounds which are chemically diverse and to produce clusters of compounds which makes sense chemically. Principal component analysis (PCA) was applied to the analysis of a small database of FT-IR spectra. The effect of the data pretreatment step of log transformation on spectral data pattern was also visualized by using PCA plots. The method of sequential projection pursuit (SPP) was applied to detect inhomogeneities in the data. Finally, cluster analysis of these spectra, depending on unweighted pair-group average linkage, was carried out.

Cluster Analysis↗

Mitochondrial control of inducible nitric oxide production in stimulated RAW 264.7 macrophages.

Addition of glucose to activated RAW 264.7 macrophages or addition of mitochondrial electron-transfer chain inhibitors enhanced the cellular nitric oxide production. An additive effect of rotenone or antimycin A and glucose on enhancing nitric oxide production was shown. Uncoupling the mitochondria by a chemical uncoupler decreased nitric oxide production. The mitochondria membrane potential was found to be important for cell viability. Although nitric oxide is the physiological inhibitor of mitochondrial respiration, this study indicates that mitochondria were not inhibited in the activated macrophages. Furthermore, a role of mitochondria in the rapid regulation of nitric oxide synthesis by the inducible nitric oxide synthase has been demonstrated.

Animals↗

A bound on mutual information for image registration.

An upper bound is derived for the mutual information between a fixed image and a deformable template containing a fixed number of gray-levels. The bound can be calculated by maximizing the entropy of the template under the constraint that the conditional entropy of the template, given the fixed image, be zero. This bound provides useful insight into the properties of mutual information as a similarity metric for deformable image registration. Specifically, it indicates that maximizing mutual information may not necessarily produce an optimal solution when the deformable transform is too flexible.

Image Processing, Computer-Assisted↗

Estradiol (E2) elicits SRC phosphorylation in the mouse neocortex: the initial event in E2 activation of the MAPK cascade?

In neocortical explants, E2 activates various signaling components of the MAPK cascade, including B-Raf and MAPK kinase-dependent ERK, suggesting a possible role in the differentiative actions of E2 in the brain. To further characterize the signaling pathways activated by E2, we determined whether c-Src, a member of the Src family of nonreceptor tyrosine kinases and an important modulator of both the MAPK cascade and neuronal differentiation, may play a role in E2 signaling. The present studies show for the first time in the brain that E2 elicits phosphorylation of c-Src on three functionally critical tyrosine residues (Y220, Y423, and Y534), and that this phosphorylation occurs despite disruption of ER alpha (in ER knockout mice). PP2, a Src family kinase inhibitor, suppressed not only E2-induced phosphorylation of c-Src, but ERK phosphorylation as well, suggesting that c-Src may be an upstream regulator of E2 signaling. E2-induced phosphorylation of c-Src is associated with increased tyrosine phosphorylation of Shc, increased association of Shc with Grb2, and induction of Ras, but not Rap1, activation. Together, these data provide evidence that E2 activates a novel c-Src-dependent signal transduction pathway in the developing brain.

Adaptor Proteins, Signal Transducing↗

[Immune response induced by hepatitis B DNA vaccine pCI-S-IRES- ProT alpha].

OBJECTIVE: To investigate immune response in mice inoculated with DNA vaccine. METHODS: pCI-S and pCI-S-IRES- ProT alpha were constructed by gene technology, transferred into cell line NIH-3T3 mediated by lipofectamine. Mice were inoculated with these plasmids too. HBsAg and HBsAb were detected by ELISA, and transcriptions of these plasmids were detected by RT-PCR. RESULTS: pCI-S -IRES- ProT alpha, and pCI-S were effectively expressed in cultured cells and vaccinated animals. Humoral immune and specific T-cell proliferative responses were stronger in pCI-S-IRES group than pCI-S group. CONCLUSIONS: Plasmids coexpression of prothymosin alpha and hepatitis B surface antigen can improve immune responses significantly.

Animals↗

[The significance of the postburn expression of proto -- oncogenes c -- fos and c -- myc mRNA and proteins in rat myocardial cells].

OBJECTIVE: To investigate the postburn expression of proto -- oncogenes c -- fos and c -- myc mRNA and proteins in rat myocardial cells following severe burn. METHODS: Wistar rats inflicted with 30% TBSA III degree burn on the back were employed as the model and randomly divided into four groups: burn (B), resuscitation (R), verapamil (V) and control (C) groups. The expressions of c -- fos and c -- myc mRNA were examined by in situ hybridization and that of the proteins by immunohistochemistry. RESULTS: The expressions of the proteins and mRNA of c -- fos and c -- myc in every experimental groups increased to peak values and then decreased along with time lapsing. And the expressions of c -- fos and c -- myc were weakest in B group, while that in V group was stronger than that in R group. CONCLUSION: The expression of postburn immediate -- early genes c -- fos and c -- myc could be induced in the myocardial cells in all the groups by severe burns. And the expression features were different from the gene expression in patients with myocardial infarction and with cardiac over -- load. This implied that there were different signal transduction and modulation mechanism among these pathological conditions.

Animals↗

[The identification of flos magnoliae by TLC].

OBJECTIVE: To establish a TLC identification method for Flos Magnoliae. METHOD: Selecting optimum TLC conditions. RESULTS: A TLC identification method of Flos Magnoliae has been established. CONCLUSION: The method is simple, accurate and reliable.

Chromatography, Thin Layer↗

[Effects of cervical vertebra rehabilitation pill on antiinflammation and analgesia].

OBJECTIVE: To study the effects of cervical vertebra rehabilitation pill on anti-inflammation and analgesia. METHODS: Hot board and body distortion induced by acertic acid methods were applied in mice for analgesia experiment. Granuloma induced by cotton ball implantation tumefaction in feet induced by carrageenin in rats and auricle tumefaction induced by croton oil in mice were applied for anti-flammation experiment. RESULTS: Cervical vertebra rehabilitation pill could significantly prolong the latent period of pain reaction induced by hot board in mice and markedly decrease the frequency of body distortion induced by acertic acid. Cervical vertebra rehabilitation pill also could significantly inhibit the tumefaction in feet induced by carrageenin, reduce the degree of tumefaction in auricle induced by croton oil in mice and inhibit the formation of granuloma induced by cotton ball implantation in subcutaneous part. CONCLUSION: Cervical vertebra rehabilitation pill has significant effects on anti-inflammation and analgesia.

Analgesics↗