The complete chloroplast genome sequence of rare and endangered Camellia pubipetala Y. Wan & S. Z. Huang (Theaceae) of South China.

The complete chloroplast genome sequence of rare and endangered Camellia pubipetala Y. Wan & S. Z. Huang (Theaceae) was mentioned in this research. By studying comparatively, we found that the C. pubipetala Y. Wan & S. Z. Huang chloroplast genome was 156,993 bp in length and composed of 86,590 bp LSC, 18,211 bp SSC, and two reverse repeating regions with 26,090 bp. The whole GC content was 37.33%. The genome encoded 116 functional genes, including 80 protein-coding genes, 32 tRNA genes, and 4 rRNA genes. In order to find the phylogenetic relationship of C. pubipetala Y. Wan & S. Z. Huang within Camellia genus, we reconstructed phylogenetic tree. The results indicate that C. pubipetala Y. Wan & S. Z. Huang was closely related to Camellia huana voucher and Camellia ptilosperma.

mTORC1 Signaling Pathway Mediates Chronic Stress-Induced Synapse Loss in the Hippocampus.

Background: The mechanistic target of rapamycin complex 1 (mTORC1) signaling has served as a promising target for therapeutic intervention of major depressive disorder (MDD), but the mTORC1 signaling underlying MDD has not been well elucidated. In the present study, we investigated whether mTORC1 signaling pathway mediates synapse loss induced by chronic stress in the hippocampus. Methods: Chronic restraint stress-induced depression-like behaviors were tested by behavior tests (sucrose preference test, forced swim test and tail suspension test). Synaptic proteins and alternations of phosphorylation levels of mTORC1 signaling-associated molecules were measured using Western blotting. In addition, mRNA changes of immediate early genes (IEGs) and glutamate receptors were measured by RT-PCR. Rapamycin was used to explore the role of mTORC1 signaling in the antidepressant effects of fluoxetine. Results: After successfully establishing the chronic restraint stress paradigm, we observed that the mRNA levels of some IEGs were significantly changed, indicating the activation of neurons and protein synthesis alterations. Then, there was a significant downregulation of glutamate receptors and postsynaptic density protein 95 at protein and mRNA levels. Additionally, synaptic fractionation assay revealed that chronic stress induced synapse loss in the dorsal and ventral hippocampus. Furthermore, these effects were associated with the mTORC1 signaling pathway-mediated protein synthesis, and subsequently the phosphorylation of associated downstream signaling targets was reduced after chronic stress. Finally, we found that intracerebroventricular infusion of rapamycin simulated depression-like behavior and also blocked the antidepressant effects of fluoxetine. Conclusion: Overall, our study suggests that mTORC1 signaling pathway plays a critical role in mediating synapse loss induced by chronic stress, and has part in the behavioral effects of antidepressant treatment.

Two new flavonoid-triterpene saponin meroterpenoids from Clinopodium chinense and their protective effects against anoxia/reoxygenation-induced apoptosis in H9c2 cells.

Two new flavonoid-triterpene saponin meroterpenoids, clinoposides G (1) and H (2) were isolated from the aerial parts of Clinopodium chinense (Benth.) O. Kuntze. Their structures were elucidated through spectroscopic and electronic circular dichroism (ECD) analyses. Compounds 1 and 2 were evaluated for their protective effects against anoxia/reoxygenation(A/R)-induced injury in H9c2 cells. A/R treatment severely injured the H9c2 cells, which was accompanied by apoptosis. Both 1 and 2 pretreatment significantly inhibited cell injury and apoptosis, improved mitochondrial membrane potential, increased activities of antioxidant enzymes, and reduced the levels of the inflammatory cytokines. In addition, the presence of 1 and 2 significantly decreased the protein level of p65 and increased the level of Nrf2 in cell nucleus. Unique chemical structure and good biological activity of 1 and 2 elucidated the potential of meroterpenoids as a promising reagent for treating heart disease.

[Studies on chemical constituents of Clinopodium chinense].

Twenty-eight compounds were isolated and purified from Clinopodium chinense by Sephedax LH-20, ODS, MCI and preparative HPLC. Their structures were identified as apigenin (1), apigenin-7-O-ß-D-glucopyranoside (2), apigenin-7-O-ß-D-glucuronopyranoside (3), thellungianol (4), apigenin-7-O-ß-D-rutinoside (5), luteolin (6), luteolin-4'-O-ß-D-glucopyranoside (7), apigenin-7-O-ß-D-pyranglycuronate butyl ester (8), luteolin-7-O-ß-D-rutinoside (9), luteolin-7-O-ß-D-noehesperidoside (10), acacetin (11), acacetin-7-O-ß-D-glucuronopyranoside (12), buddleoside (13), naringenin (14), pruning (15), nairutin (16), isosakuranetin (17), isosakuranin (18), didymin (19), hesperidin (20), kaempferol (21), quercetin (22), kaempferol-3-O-α-L-rahmnoside (23), p-hydroxycinnamic acid (24), caffeic acid (25), cis-3-[2-[1-(3,4-dihydroxy-phenyl)-1 -hydroxymethyl]-1,3-ben-zodioxol-5-yl]-(E)-2-propenoic acid (26), mesaconic acid (27), gentisic acid 5-O-ß-D-(6'-salicylyl)-glucopyranoside (28). Among them, compounds 7, 9-10, 12, 23, 26-28 were isolated from the Clinopodium for the first time. The protective effects of compounds 1-6, 8-17 and 19 against H2O2-induced H9c2 cardiomyocyte injury were tested, compounds 15 exhibited significantly protective effects. Compared with the cell viability of (62.12±6.18)% in the model, pruning exhibited viabilities of (84.25±7.36)% at 25.0 mg•L⁻¹, respectively, using quercetin as a positive control [cell viability of (84.55±8.26)%, 20 mg•L⁻¹].

Two new tetracyclic triterpenoids from the barks of Melia azedarach.

Two new tetracyclic triterpenoids, together with 21 known compounds, were isolated from the barks of Melia azedarach. The structures of new compounds were elucidated by the means of HRESIMS, 1D NMR, 2D NMR, and X-ray crystallography analysis.

Crystal structure of 16-hy-droxy-4,4,10,13,14-penta-methyl-17-(6-methyl-hept-5-en-2-yl)-4,5,6,9,10,11,12,13,14,15,16,17-dodeca-hydro-1H-cyclo-penta-[a]phenanthren-3(2H)-one.

The title compound, C30H48O2, contains a fused four-ring triterpenoid system. In the mol-ecule, the two cyclo-hexane rings adopt a chair conformation and a twist boat conformation, respectively, the central cyclo-hexene ring adopts a half-chair conformation whereas the five membered ring adopts an envelope conformation. In the crystal, O-H⋯O hydrogen bonds between the hy-droxy and carbonyl groups of adjacent mol-ecules link the mol-ecules into supra-molecular chains propagating along the b-axis direction.

Quantification of the sixth DNA base 5-hydroxymethylcytosine in colorectal cancer tissue and C-26 cell line.

BACKGROUND: DNA methylation at the five position of cytosine is well recognized as an important epigenetic modification in human health and disease. Recent evidences demonstrated that 5-methylcytosine (5-mC) by the TET family of enzymes can be converted to 5-hydroxymethylcytosine (5-hmC). Here, we use an ultrasensitive and accurate isotope-based LC-MS/MS method to precisely determine the levels of 5-hmC and 5-mC in colorectal cancer and the C-26 colon adenocarcinoma cell line. RESULTS: Our data showed that 5-hmC content is significantly reduced (approximately sixfold) in colorectal cancer as compared with adjacent normal tissue. Similarly, the ratio of 5-hmC to 5-mC dropped from 0.054 ± 0.005 in normal tissues, to 0.011 ± 0.002 in cancer. CONCLUSION: The analysis of 5-hmC levels and the ratio of 5-hmC:5-mC during tumor progression might provide insight into the role of this modification in cellular immortalization and transformation.

Synthesis and biological evaluation of some novel Schiff's bases from 1,2,4-triazole.

We have synthesized a number of novel Schiff's bases from 4-amino-3-(D-glucoheptonic-hexitol-1-yl)-1H-1,2,4-triazole-5-thione. By attaching D-glucoheptonic-hexitol-1-yl residues to 1,2,4-triazole at the 3-position, the solubility of the title compounds has been improved greatly. All the products have been characterized by elemental analysis, IR, 1H-NMR, 13C-NMR and MS. The plant-growth regulating effects of the title compounds were examined. From the biological activity results, we found that most compounds showed weak to moderate activities.

P5644 interacts with phosphatidylinositol-4-phosphate adaptor protein-1 associated protein-1.

The human novel gene pp5644 (GeneBank Accession No. AF289559) coding for 124 amino acids was recently cloned. Overexpression of pp5644 in Hela cells significantly inhibited the growth and colony formation. The pp5644-interacting protein FAPP1 (phosphatidylinositol-four-phosphate adaptor protein1) associated protein-1, called FASP1, was obtained by using yeast two-hybrid system. The interaction between pp5644 and FASP1 was experimentally confirmed by GST pull-down assay in vitro and co-immunoprecipitation assay in vivo. Co-localization of pp5644 and FASP1 in cytoplasm in Hela cells could further support the interaction. Based on the experimental results, it is suggested that pp5644 physically bind to FASP1 and the biological significance of this kind of interaction in vivo is discussed.

[Enantiomeric separation of ketoprofen by HPLC using chirobiotic V CSP and vancomycin as chiral mobile phase additives].

AIM: To establish HPLC chiral separation method for ketoprofen enantiomers by using Chirobiotic V chiral seperation phase (CSP) (A) and vancomycin as chiral mobile phase additives (B). METHODS: The separation was first performed on Chirobiotic V CSP with the mobile phase of terahydrofran (THF)-0.5% triethylanine acetate(TEAA) buffer (15:85) at the flow rate of 0.7 mL.min-1. When using vancomycin as chiral mobile phase additive, the separation was carried out on C8 column (150 mm x 4.6 mm), the mobile phase was methanol-0.25% TEAA buffer (50:50), the flow rate was 0.7 mL.min-1. The effects of the concentration of vancomycin, organic modifier and the pH of the buffer on the resolution of ketoprofen enantiomers were investigated. Also, the feasibility of these two methods to be used as quantitative method was studied. RESULTS: Ketoprofen enantiomers were separated at a baseline level under the chromatographic condition of both methods A and B, the resolution was 2.28 and 2.22, respectively. In method A the linearity of enantiomer was obtained from 0.5 mg.L-1 to 100 mg.L-1, the detectionlimit was 1 microgram.L-1. When using vancomycin as mobile phase additive the system was shown to have a high efficiency. In this system, the assay of enantiomer is linear from 2.5 mg.L-1 to 250 mg.L-1. The detection limit was 14.5 micrograms.L-1. CONCLUSION: Both methods can be used to detect optical purity of S-(+)-ketoprofen.