Effect of cytochrome P450 3A4 on tissue distribution of humantenmine, koumine, and gelsemine, three alkaloids from the toxic plant Gelsemium.

Humantenmine, koumine, and gelsemine are three indole alkaloids found in the highly toxic plant Gelsemium. Humantenmine was the most toxic, followed by gelsemine and koumine. The aim of this study was to investigate and analyze the effects of these three substances on tissue distribution and toxicity in mice pretreated with the Cytochrome P450 3A4 (CYP3A4) inducer ketoconazole and the inhibitor rifampicin. The in vivo test results showed that the three alkaloids were absorbed rapidly and had the ability to penetrate the blood-brain barrier. At 5 min after intraperitoneal injection, the three alkaloids were widely distributed in various tissues and organs, the spleen and pancreas were the most distributed, and the content of all tissues decreased significantly at 20 min. Induction or inhibition of CYP3A4 in vivo can regulate the distribution and elimination effects of the three alkaloids in various tissues and organs. Additionally, induction of CYP3A4 can reduce the toxicity of humantenmine, and vice versa. Changes in CYP3A4 levels may account for the difference in toxicity of humantenmine. These findings provide a reliable and detailed dataset for drug interactions, tissue distribution, and toxicity studies of Gelsemium alkaloids.

Antinociceptive effect of gelsenicine, principal toxic alkaloids of gelsemium, on prostaglandin E2-induced hyperalgesia in mice: Comparison with gelsemine and koumine.

Gelsemium elegans (G.elegans) is a plant of the Loganiaceae family, known for its indole alkaloids, including gelsemine, koumine, and gelsenicine. Gelsemine and koumine are well-studied active alkaloids with low toxicity, valued for their anti-anxiety and analgesic properties. However, gelsenicine, another important alkaloid, remains underexplored due to its high toxicity. This study focuses on evaluating the analgesic properties of gelsenicine and comparing them with gelsemine and koumine. The results indicate that all three alkaloids exhibit robust analgesic properties, with gelsemine, koumine, and gelsenicine showing ED50 values of 0.82 mg/kg, 0.60 mg/kg, and 8.43 µg/kg, respectively, as assessed by the hot plate method. Notably, the therapeutic dose of gelsenicine was significantly lower than its toxic dose (LD50 = 0.185 mg/kg). The study also investigated the mechanism of action by analyzing the expression levels of GlyRα3 and Gephyrin. The PGE2 model group showed decreased expression levels of GlyRα3 and Gephyrin, while groups treated with gelsemine, koumine, and gelsenicine were able to reverse this decrease. These results suggest that gelsenicine effectively alleviates PGE2-induced hyperalgesia by upregulating the expression of GlyRα3 and Gephyrin, which are key targets of the Gly receptor pathway.

Effect of heavy metal stress on emerging plants community constructions in wetland.

In this paper, we report the effects of heavy metals (HMs) (cadmium and mercury) on seed germination and seedling growth of Phragmites australis and Triarrhena sacchariflora, which are the two main typical emerging plants in Hongze Lake wetland. The results showed that there was a reduction in germination percentage, germination index and seedling length as HM concentration in the growing media increased for both treatments. The effect of HMs toxicity on seed germination and seedling growth of T. sacchariflora was more obvious than of P. australis. At the stage of seed germination, P. australis and T. sacchariflora were sensitive to Hg(2 + ) and Cd(2 + ), respectively, and Hg(2 + ) was more toxic than Cd(2 + ) at the stage of seedling growth. The effect of HMs toxicity is not invariable during plant growth. Compared to the stage of seedling growth, P. australis and T. sacchariflora are more susceptible to HMs at the stage of seed germination. In addition, we calculated the ecological thresholds of P. australis to Cd and Hg are 19.32 and 1.08 mg kg(-1), and that of T. sacchariflora to Cd is 4.62 mg kg(-1) based on the lab simulation. The results also indicated that the species of P. australis is more tolerant than T. sacchariflora to the HMs and is a better candidate for restoration in Hongze Lake wetland ecosystem.

Optimal Supply of Precursors for CoQ10 Production by Rhodopseudomonas Palustris / 现代生物医学进展

CoQ10 has been used not only as a medicine but also as food supplements because of its various physiological and biochemical activities. A full-factorial central composite design and response surface methodology were used for optimizing three precursors Solanesol, 4-hydroxybenzoic acid and methionine to maximize the production of CoQ10 by Rhodopseudomonas palustris J001. The optimization of the model predicted a maximum response 40.6 [(mg CoQ10)(g dried biomass)-1] CoQ10 production with 124.8 mg l-1 Solanesol, 267.7 mg l-14-hydroxybenzoic acid and 130.2 mg l-1 methionine, respectively. A new combination was prepared according to the result. The observed response was 40.5 ± 0.2 [(mg CoQ10)(g dried biomass)-1] and was 109.8%higher than in the control with no addition of the three precursors.

Optimal Supply of Precursors for CoQ10 Production by Rhodopseudomonas Palustris / 现代生物医学进展

CoQ10 has been used not only as a medicine but also as food supplements because of its various physiological and biochemical activities. A full-factorial central composite design and response surface methodology were used for optimizing three precursors Solanesol, 4-hydroxybenzoic acid and methionine to maximize the production of CoQ10 by Rhodopseudomonas palustris J001. The optimization of the model predicted a maximum response 40.6 [(mg CoQ10)(g dried biomass)-1] CoQ10 production with 124.8 mg l-1 Solanesol, 267.7 mg l-14-hydroxybenzoic acid and 130.2 mg l-1 methionine, respectively. A new combination was prepared according to the result. The observed response was 40.5 ± 0.2 [(mg CoQ10)(g dried biomass)-1] and was 109.8%higher than in the control with no addition of the three precursors.

Studies on the protective immunity of Schistosoma japonicum bivalent DNA vaccine encoding Sj23 and Sj14.

In order to explore the high performance bivalent DNA vaccine of Schistosoma japonicum, the fatty-acid-binding protein (Sj14) and the 23 kDa transmembrane protein (Sj23) two proteins were selected to construct the DNA-based vaccine. It was successful to construct a bivalent DNA vaccine using three strategies: the co-expression of two genes, a fusion gene expression and two kinds of plasmids in combination (cocktail vaccine). The bivalent DNA was proven to express well in vitro and in vivo by indirect immunofluorescence test (IIF) and reverse transcriptase-polymerase chain reaction (RT-PCR). The protective immunity of bivalent DNA vaccine was higher than that of univalent DNA vaccine (p<0.05). There were four groups of bivalent vaccine whose protective immunity was higher than 50%. Granuloma diameter reduction rates were in the range of 18-39%. There was no significant impact on immunity protection exerted by the four factors including dosage, inoculated times, inoculated routes and challenge time after the last immunization in three levels (p>0.05).

Analyzing Role of △~5 Desaturase in Biosynthesis of Arachidonic Acid Using Real Time PCR / 中国生物工程杂志

Arachidonic acid is an essential fatty acid in human nutrition and a biogenetic precursor of the biologically active prostaglandins and leukotrienes. ?~5 desaturase catalyzes the ?~5 dehydrogenation of di-homo-?-linolenic acid to form arachidonic acid in biosynthetic pathway of arachidonic acid. Using real time PCR technology,the transcriptional expression levels of gene encoding ?5 desaturase in three Mortierella alpina strains M10,M6 and M23,and in different growth phase of high arachidonic acid yielding strain M6,were determined. Results showed that there was a distinct corelationship between mRNA transcript level of ?~5 desaturase gene and biosynthesis of arachidonic acid. Results indicated that ?~5 desaturase plays an important role in arachidonic acid biosynthesis.

Culture Conditions Optimization of Magnetotactic Bacteria WM-1 / 现代生物医学进展

The culture of Magnetospirillum magneticum WM-1 depends on several control factors that have great effect on the magnetic cells concentration. Investigation into the optimal culture conditions needs a large number of experiments. So it is desirable to minimize the number of experiments and maximize the information gained from them. The orthogonal design of experiments and mathematical statistical method are considered as effective methods to optimize the culture condition of magnetotactic bacteria WM-1 for high magnetic cells concentration. The effects of the four factors, such as pH value of medium, oxygen concentration of gas phase in the serum bottle, C:C (mtartaric acid: msuccinic acid) ratio and NaNO3 concentration, are simultaneously investigated by only sixteen experiments through the orthogonal design L16(44) method. The optimal culture condition is obtained. At the optimal culture condition (pH 7.0, a oxygen concentration 4.0%, C: C (mtartaric acid: msuccinic acid) ratio 1:2 and NaNO3 100 mg l-1), the magnetic cells concentration is promoted to 6.5×107 cells ml-1, approximately 8.3% higher than that under the initial conditions. The pH value of medium is very important factor for magnetic cells concentration. It can be proved that the orthogonal design of experiment is of 90% confidence. The results from the hysteresis of WM-1 shows that Hc = 230 Oe, Ms = 0.9 emu/g dry wt. Cells,and Mr / Ms = 0.50.

Culture Conditions Optimization of Magnetotactic Bacteria WM-1 / 现代生物医学进展

The culture of Magnetospirillum magneticum WM-1 depends on several control factors that have great effect on the magnetic cells concentration. Investigation into the optimal culture conditions needs a large number of experiments. So it is desirable to minimize the number of experiments and maximize the information gained from them. The orthogonal design of experiments and mathematical statistical method are considered as effective methods to optimize the culture condition of magnetotactic bacteria WM-1 for high magnetic cells concentration. The effects of the four factors, such as pH value of medium, oxygen concentration of gas phase in the serum bottle, C:C (mtartaric acid: msuccinic acid) ratio and NaNO3 concentration, are simultaneously investigated by only sixteen experiments through the orthogonal design L16(44) method. The optimal culture condition is obtained. At the optimal culture condition (pH 7.0, a oxygen concentration 4.0%, C: C (mtartaric acid: msuccinic acid) ratio 1:2 and NaNO3 100 mg l-1), the magnetic cells concentration is promoted to 6.5×107 cells ml-1, approximately 8.3% higher than that under the initial conditions. The pH value of medium is very important factor for magnetic cells concentration. It can be proved that the orthogonal design of experiment is of 90% confidence. The results from the hysteresis of WM-1 shows that Hc = 230 Oe, Ms = 0.9 emu/g dry wt. Cells,and Mr / Ms = 0.50.

LC-ESI-MS metabolic profiling analysis of taxanes from the extracts of Taxus chinensis cell cultures / 药学学报

<p><b>AIM</b>To develop a rapid analytical method for small amount biological samples of taxanes by using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS) in small amount biological samples.</p><p><b>METHODS</b>A solution containing five given taxane constituents and the extract from cell cultures of Taxus chinensis were analysed separately. According to the performance of the given taxanes in ESI-MS/MS, run parameters of the mass spectrometer were optimized. Positive and negative electrospray modes were employed to simultaneously scan the cell cultures sample, and the full ion chromatogram and the molecular weight of individual peak were obtained. The qualitative analysis of taxanes was achieved by comparison of the retention time and molecular weight with those of the reference substances or was based on the interpretation of the MS/MS spectra of the analytes and the knowledge of the concerning genetic backgrounds of taxanes published in literatures.</p><p><b>RESULTS</b>The taxanes with several acetyl substituents tended to produce ammonium adduct ions peak, while multi-hydroxy taxanes were subject to give protonized molecular ion peaks in positive ion ESI-MS. Thirteen taxanes in cell samples were assigned. Eight compounds of them were identified to be 1 -acetyl-5, 7, 10-deacetyl-baccatin I (DAB-I, 1) , baccatin III (B-III, 3), baccatin VI (B-VI, 8), taxol (9), yunnanxane (10 ), taxuyunnanine C (Tc, 11), sinenxane B (12), sinenxane C (13), separately. For the other five constituents, character of taxane and the number of substituents were deduced.</p><p><b>CONCLUSION</b>The results confirm the feasibility of characterizing taxanes in biological samples by LC-ESI-MS analysis. The analytical methodology provided a rapid, conventional and reliable tool to study metabolic profiling of taxanes for structural elucidation in taxol biosynthesis.</p>

Cloning and analysis of cDNA encoding key enzyme gene (dxr) of the non-MVA pathway in Taxus chinensis cells / 生物工程学报

Two distinct routes (classical mevalonate pathway and a novel mevalonate-independent pathway) are utilized by plants for the biosynthesis of isopentenyl diphosphate, the universal precursor of isoprenoids (Fig. 1). Present researches indicated that taxol was synthesized mainly via non-mevalonate pathway, but not genetic evidence was showed. The second step in non-mevalonate pathway involves an intramolecular rearrangement and subsequent reduction of deoxyxylulose phosphate to yield 2-C-methyl-D-erythritol-4-phosphate, and 1-Deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) with responsibility for this reaction was considered as a key enzyme. As a tool for the isolation of genes in terpenoid biosynthesis in plants, total RNA was prepared from Taxus chinensis suspension cells, a cell type highly specialized for diterpene (taxol). A reverse transcription-PCR strategy based on the design of degenerated oligonucleotides was developed for isolating the gene encoding a gymnosperm homolog of this enzyme from Taxus chinensis. Through sequence analysis by Blast P online, the resulting cDNA showed highly homologous to 1-deoxy-D-xylulose 5-phosphate reductoisomerases, with 95% identification compared with Arabidopsis thaliana (Q9XFS9), 94% with Mentha x piperita (Q9XESO), 80% with Synechococcus elongatus (Q8DK30), 78% with Synechocystis sp. PCC 6803 (Q55663) and Nostoc sp. PCC 7120 (Q8YP49), and 73% with Synechococcus leopoliensis (Q9RKT1). Deduced amino acid sequences were also analyzed by PROSITE, ClustalX (1.81) and Phylio (3.6 alpha), and data present evidence for the existence of this deoxyxyluose phosphate reductoisomerase in Taxus chinensis. This is the first report of the dxr gene cloned from gymnosperm.

Effects of Karst Environmental Factors on Expression and Activity of Bacterial Extracellular Carbonic Anhydrase / 微生物学通报

Effects of main environmental factors, such as temperature, pH , metal ions and anions, on expression and activity of bacterial extracellular car bonic anhydrase(CA) were studied, exemplified by a bacterium named GLRT102Ca, wh ich was separated from karst ecosystems of Southwest China. The results showed that the tested strain could express different activity of extracellular carboni c anhydrase within the scope of experimental temperature (10℃~50℃) and pH (5. 5~9.0). The activity of extracellular carbonic anhydrase was higher at tempera ture of 20℃~30℃ and at neutral and alkaline trending condition. Moreover, the expression of activity of extracellular carbonic anhydrase could be generally p romoted at the experimental range of concentration of 4 kinds of metal ions such as Ca2+, Mg2+, Zn2+ and Co2+, along with 8 kinds of ani ons such as SO2-_4, H_2PO-_4, NO-_3, NO-_2, Cl-,Br-, I- and HCO-_3. This research provides a cert ain theoretical basis for further study on the role of microbial CA in karst pro cesses.