Objective To investigate the effects and mechanisms of all-trans retinoic acid(ATRA)on the proliferation and cell cycle of lung carcinoma cell line A549.Methods The A549 cells were treated with ATRA at the dosages of 5,10,50 ?mol/L for 1-7 d.The proliferation of A549 was assessed by MTT method and cell cycle was analyzed by flow cytometry.The expressions of CDK4,Rb and p-ERK1/2 were assessed by Western blotting.CyclinD1 mRNA was analyzed by SYBR-PCR amplification.Results ATRA obviously inhibited the proliferation of A549 cells,and the cell cycle was arrested in G0/G1 phase.The expression of p-ERK1/2 protein and CyclinD1 mRNA on A549 cells were decreased.Conclusion ATRA might inhibit the proliferation of A549 cells through down-regulating p-ERK1/2 protein and CyclinD1 mRNA.

Aortic Aneurysm ; diagnosis ; Aortic Aneurysm, Thoracic ; diagnosis ; Carotid Body Tumor ; diagnosis ; Diagnostic Errors ; Humans ; Male ; Middle Aged

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

Cooperation hydrogen production was carried out using Rhodopseudomonas sp. DT and Enterobacter aerogenes. The effects of the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes, culture temperature, and carbon source on the cooperation hydrogen production were investigated. The results suggested that cooperation hydrogen production rate was highly affected by the initial ratio of Rhodopseudomonas sp. DT and E. aerogenes. The mixed bacteria of Rhodopseudomonas sp. DT and E. aerogenes with 1:1 initial ratio benefited to the cooperation hydrogen production, which led the hydrogen production rate and duration of gas production to 3.1 mol H2/mol glucose and 81 h, respectively. The pH dynamics analysis of culture medium further discovered that the pH of the mixed bacteria with 1:1 initial ratio changed from 6 to 7 smaller than other conditions, which was probably fitted to produce hydrogen. Furthermore, the mixed bacteria with 1:1 initial ratio had the higher hydrogen production efficiency at temperatures of 28?C and 37?C than at 20?C, and without any hydrogen production at temperature of 50?C. The carbon sources of glucose, succinate acid, malic acid could be used to produce hydrogen by the mixed bacteria. Even the soluble starch, unused by Rhodopseudomonas sp. DT, was also decomposed by the mixed bacteria to produce hydrogen with the conversion efficiency of 8.22%. The glucose was the optimal carbon resource, and the conversion efficiency could reach to 36.11%. The results, further, implied that the cooperation hydrogen production could enlarge the use of the carbon sources.

Objective To investigate the relatonahip between TNF-α and ventricular arrhythmias after acute myocardial infarction(AMI)and its mechanism.Method Both the clinical and animal experiments were done.(1)Clinical experiment:Eighty patients with AMI were included in Union Hospital,Tongji Medical College,Huazhong University of Science and Techology,from May 2005 to November 2006 according to the WHO diagnostic criteria.Co-infection of diseases such as severe upper respiratory infection,lung infection,high fever,cancer,et al were excluded.The relationship between the levels of TNF-α and arrhythmias were observed at different times after AMI.A straight line correlation,analysis Was done.(2)Animal experiment:Different concentrations of TNF-αwere added to isohted rat hearts for observing the arrhythrnia effects.The effect of TNF-α on intracellular Ca2+ concentration was detected by laser confocal technique.All data were analyzed by SNK-q test using SPSS 13.0 sofeware prograrn.Results(1)The plasma levels of TNF-α were significantly associated with the Lown class of PVC after AMI and they were higher in AMI of anterior wall[(46.41±10.34)pg/ml]than other positions [(28.25±6.35)pg/ml,P<0.05].2)The frequency of ventricular arrhythmias was interrelated with the concentralions of TNF-α.Using etanercept beforehand,TNF-α induced a slight increase of intracellular Ca2+ intensity (P<0.05).Conclusions There was a relationship between TNF-αlevels and ventricular arrhythmisa in patients with AMI.Animal experiments confirmed the isolated heart perfusion with TNF-α induced ventricular arrhytrnias.Expression of TNF-α after AMI was related with the occurrence of ventricular arrhythrnias.The effect might be associated with the increased inuaeellular Ca2+ intensity caused by TNF-α.

Cochlear Nerve ; chemistry ; pathology ; Cranial Nerve Neoplasms ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Heart Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Neurilemmoma ; metabolism ; pathology ; S100 Proteins ; metabolism ; Vestibulocochlear Nerve Diseases ; metabolism ; pathology ; Vimentin ; metabolism

Referring to the rules for agricultural seed testing (GB /T 3543-1995) issued by China, the test of sampling, seed purity, weight per 1 000 seeds, seed moisture, seed viability and germination rate had been studied for screening seed quality test methods of Pesudostellaria heterophylla. The seed quality from different collection areas was measured. The results showed that at least 6.5 g seeds should be sampled and passed through 10-mesh sieve for purity analysis. The weight of 1 000 seeds was determined by using the 500-seed method. The phenotypic observation and size measurement were used for authenticity testing. The seed moisture was determined under the higher temperature (130 ± 2) degrees C for 5 hours. The seeds were dipped into 0.2% TTC sustaining 1 hour at 40 degrees C, then the viability could be determined. The break dormancy seeds were cultured on sand at 10 degrees C. K cluster analysis was applied for the data analysis, the seed quality from different collection areas grading of P. Heterophylla was described as three grades. The seed quality of each grade should reach following requirements: for first grade seeds, germination rate ≥ 86%, 1 000-grain weight ≥ 2.59 g, purity ≥ 87%, moisture ≤ 13.1%; for second grade seeds, germination rate ≥ 70%, 1 000-grain weight ≥ 2.40 g, purity ≥ 77%, moisture ≤ 14.3%; for third grade seeds, germination rate ≥ 41%, 1 000-grain weight ≥ 2.29 g, purity ≥ 76%, moisture ≤ 15.8%. The seed testing methods for quality items of P. heterophylla had been initially established, as well as the primary P. heterophylla seed quality classification standard.
Caryophyllaceae ; chemistry ; growth & development ; Germination ; Quality Control ; Seeds ; chemistry ; growth & development ; Water ; analysis

Objective To conduct mutation screening of SCN1A 3′ untranslated region (UTR) on Dravet syndrome (DS) patients without mutations in the SCN1A coding region and promoter region, and functional analysis of the variant from DS patients.Methods Twenty-eight DS patients without mutations in the SCN1A coding region and promoter region were screened for SCN1A 3′ UTR mutations using PCR and direct sequencing.Functional analysis of the detected mutation was done via luciferase assay, mRNA stability analysis and RNA electrophoretic mobility shift assay (RNA-EMSA).Results A novo variant (c.*20A>G) in SCN1A 3′ UTR was found in one DS patient.The variant (c.*20A>G) reduced the luciferase gene xpression by 30% through increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing luciferase gene mRNA stability (t=8.5,P<0.01).Conclusions A functional variant was detected from one patient with DS.This variant negatively regulated the gene expression by increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing mRNA stability.

Objective To investigate cyclic mechanical stimulation on expression of connective tissue growth factor (CTGF) in osteoblast-like cells (MG63) and to explore the rote of MAPK involved in the process.Methods Expressions of CTGF protein and mRNA in MG63 cells were detected by Western blot and RT-PCR,respectively. Phosphorylation levels of p38, ERK, JNK were examined by Western blot. Results Cyclic mechanical stimulation upregulated expressions of CTGF protein and mRNA. The levels reached a maximal response of 2-3 fold after 3-6 h. ERK and JNK signal pathways were activated by cyclic mechanical stimulation, the phosphorylated proteins increased within 10 min of stretch, phosphorylated ERK reached maximal levels by 60 min of stretch, phosphorylated JNK reached maximal levels by 15-30 min of stretch, but not for p38 signal pathway.Only the inhibitior of JNK signal pathway (SP600125) markedly suppressed stretch-induced CTGF expression,meanwhile the inhibitors of ERK (PD98059) and p38 (SB203580) did not show such effect. Conclusion Cyclic mechanical stimulation upregulates CTGF expression via JNK-dependent pathway in MG63 cells.

This dissertation is to determine the biopotency of hemostat which processed in different places by establishing a bioassay method of Bletillae Rhizoma based on the thrombin time. Contrast test is the main methodology. Specifically, the reference substance of Bletillae Rhizoma is determined by comparing with the control substance of vitamin K1 using thrombin time, which is calibrated the Bletillae Rhizoma. The hemostatic biopotency is calculated by using the method of "parallel line assay method based on quantitative responses" (3.3) from different processed products. It indicates that there is a strong linear correlation between Bletillae Rhizoma and control drugs (Y = 66.332-23.913X, R2 = 0.995 3). The hemostatic biopotency of Bletillae Rhizoma from different processed products ranged between 821.93-1 187.53 U x g(-1) shown in the paper, and all of them can meet the requirements of the test. The methodology has an appropriate instrument precision (RSD 3.8%), intermediate precision (RSD 4.6%), repeatability (RSD 3.2%) and stability (RSD 3.7%). Therefore, it can be turned out that the methodology which established in the dissertation is good at determinating the hemostatic biopotency of Bletillae Rhizoma and it is reliable, simple and repeatable.
Animals ; Drugs, Chinese Herbal ; pharmacology ; standards ; Hemostatics ; pharmacology ; standards ; Orchidaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Thrombin Time