AIM: To investigate the ornithine decarboxylase (ODC) gene expression in colorectal carcinoma, ODC mRNA was assayed by RT-PCR and ODC protein was detected by a monoclonal antibody against fusion of ...human colon ODC prepared by hybridoma technology.METHODS: Total RNA was extracted from human colorectal cancer tissues and their normal counterpart tissues. ODC mRNA levels were examined by RT-PCR.ODC genes amplified from RT-PCR were cloned into a prokaryotic vector pQE-30. The expressed proteins were purified by chromatography. Anti-ODC mAb was prepared with classical hybridoma techniques and used to determine the ODC expression in colon cancer tissues by immunohistochemical and Western blotting assay.RESULTS: A cell line, which could steadily secrete antiODC mAb, was selected through subcloning four times. Western blotting reconfirmed the mAb and ELISA showed that its subtype was IgG2a. RT-PCR showed that the ODC mRNA level increased greatly in colon cancer tissues (P<0.01). Immunohistochemical staining showed that colorectal carcinoma cells expressed a significantly higher level of ODC than normal colorectal mucosa (98.6±1.03% vs 5.26±5%, P<0.01).CONCLUSION: ODC gene overexpression is significantly related to human colorectal carcinoma. ODC gene expression may be a marker for the gene diagnosis and therapy of colorectal carcinoma.
Brain‐derived neurotrophic factor (BDNF) has been implicated in the potent modulation of synaptic plasticity at both pre‐synaptic and post‐synaptic sites. However, the molecular mechanism underlying ...BDNF‐mediated pre‐synaptic modulation remains incompletely understood. Here, we report that BDNF treatment for over 4 h could significantly enhance the expression of c‐Jun NH2‐terminal kinase‐interacting protein 3 (JIP3) in cultured hippocampal neurons. This enhancement could be blocked by the Trk inhibitor K252a or by a cAMP response element‐binding protein (CREB) inhibitor. In addition, chromatin immunoprecipitation (ChIP) assays revealed that CREB could bind with the JIP3 promoter region and the BDNF treatment could increase this binding. Using dual‐luciferase assays we further characterized the cAMP response element (CRE) site in the JIP3 promoter. Finally, we found that BDNF‐increased JIP3 expression contributes to the BDNF‐induced modulation of neurotransmitter release. Together, our studies reveal that in hippocampal neurons BDNF up‐regulates JIP3 expression via CREB activation, which contributes to the enhancement of neurotransmitter release; thus, we have identified a novel mechanism that BDNF modulates pre‐synaptic transmission.
We demonstrated that in hippocampal neurons BDNF/TrkB signaling mediates transcriptional up‐regulation of c‐Jun NH2‐terminal kinase‐interacting protein 3 (JIP3) via CREB activation. The up‐regulation of JIP3 further contributes to BDNF‐enhanced neurotransmitter release. These findings provide insight into the mechanistic link between BDNF‐mediated gene expression and its more sustained pre‐synaptic modulation, which may help us to further understand the roles of BDNF in neuronal plasticity.
We demonstrated that in hippocampal neurons BDNF/TrkB signaling mediates transcriptional up‐regulation of c‐Jun NH2‐terminal kinase‐interacting protein 3 (JIP3) via CREB activation. The up‐regulation of JIP3 further contributes to BDNF‐enhanced neurotransmitter release. These findings provide insight into the mechanistic link between BDNF‐mediated gene expression and its more sustained pre‐synaptic modulation, which may help us to further understand the roles of BDNF in neuronal plasticity.
The possibility of determination of tofu hardness by near infrared spectroscopy (NIR, 833-2500 nm) was studied. The influences of water content and protein content on the spectra were discussed, in ...order to detect the hardness. The models of Tofu hardness were calibrated by partial least squares regression (PLS) after eliminating outline, spectra pretreatment, and wavelength optimization (r = 0.994, RMSEC = 0.391%, RMSEP = 0.416%, RSD = 2.27%). All of the results were better than those by sense assessing method.
Studying the essence of a syndrome has been a key challenge in the field of Chinese medicine.Until now,due to limitations of the methods available,the progress towards understanding such complicated ...systems has been slow.Metabonomics encompasses the dynamics,composition and analysis of metabolites,enabling the observation of changes in the metabolic network of the human body associated with disease.Being from the point of view of the whole organism,metabonomics provides an opportunity to study the essence of a syndrome to an unprecedented level.Phlegm and blood stasis syndrome is the main syndrome associated with coronary heart disease（CHD）,which bring difficulties in clinical treatment due to difficulties associated with differentiation of symptoms and signs.The fundamental differences of material between the two also need to be interpreted.The authors consider that we can use the method of combining a disease（in this case CHD）with associated syndromes（phlegm and blood stasis syndrome）to select patients with phlegm and blood stasis syndrome of CHD,and utilize metabonomics to explore the essence of the syndrome by difference analysis of metabolite spectra.Meanwhile,we can study the syndrome in CM,observe the change regularity of metabolism spectra after the treatment of corresponding and non-corresponding prescription and syndrome,in order to validate the material fundament in the progress of syndrome formation and their differences.This will not only have great significance in enhancing the ability to identify syndrome of phlegm and blood stasis in CHD and to establish the clinical curative criteria,but will also offer a new approach of studying the essence for a syndrome using metabonomics.
Two novel bioactive mono-tetrahydrofuran (THF) γ-lactone acetogenins, 16,19-
cis-murisolin (
1) and murisolin A (
2), were isolated from the seeds of
Asimina triloba (Annonaceae) by directing the ...fractionation with brine shrimp lethality.
1 has a 16,19-
cis mono-THF ring with two flanking hydroxyl groups and represents a new type among the natural Annonaceous acetogenins. Murisolin (
3), which is new in this species, was also obtained.
1–3 showed potent and selective cytotoxicities among six human tumor cell lines.
Two new Annonaceous acetogenins, 16,19-
cis-murisolin (
1) and murisolin A (
2), were isolated from the seeds of
Asimina triloba.
To construct and express ODC recombinant gene for further exploring its potential use in early diagnosis of colorectal carcinoma.
Total RNA was extracted from colon cancer tissues and amplified by ...reverse-transcription PCR with two primers, which span the whole coding region of ODC. The synthesized ODC cDNA was cloned into vector pQE-30 at restriction sites BamH I and Sal I which constituted recombinant expression plasmid pQE30-ODC. The sequence of inserted fragment was confirmed by DNA sequencing, the fusion protein including 6His-tag was facilitated for purification by Ni-NTA chromatographic column.
ODC expression vector was constructed and confirmed with restriction enzyme digestion and subsequent DNA sequencing. The DNA sequence matching on NCBI Blast showed 99 % affinity. The vector was transformed into E. coli M15 and expressed. The expressed ODC protein was verified with Western blotting.
The ODC prokaryote expression vector is constructed and thus greatly facilitates to study the role of ODC in colorectal carcinoma.
A novel acetogenin, asiminocin (1), was isolated by activity-directed fractionation from the stem bark of the paw paw tree,
Asimina triloba. By spectral and chemical methods, 1 was identified as (30
...S)-hydroxy-4-deoxyasimicin. The absolute configuration of 1, along with those of previously reported acetogenins asimin, asiminacin, bullatin, (30
S)-bullanin, and (30
R) bullanin, was determined by Mosher ester methodology. Compound 1 was highly inhibitory to three human solid tumor cell lines with over a billion times the potency of adriamycin.
Asiminocin (1), a novel acetogenin isolated from
Asimina triloba, was identified as 30
S-hydroxy-4-deoxyasimicin. Compound 1 was highly inhibitory to three human solid tumor cell lines with over a billion times the potency of adriamycin.
The leaves of Annona muricata have yielded eight monotetrahydrofuran Annonaceous acetogenins. Two of them, annomuricins A 1 and B 2, whose chemical structures were deduced by ms, nmr, ir, and uv ...spectral and chemical methods, are novel and unusual. Compounds 1 and 2 each possess five hydroxyl groups; two hydroxyl groups are vicinal, with the vicinal group of 1 threo and that of 2 erythro. The absolute configurations of 1 and 2 were determined by Mosher ester methodology. Six monotetrahydrofuran acetogenins, previously described in the seeds, were found in the leaves; these are gigantetrocin A, annonacin-10-one, muricatetrocins A and B, annonacin, and goniothalamicin.
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