CgPG21 is involved in the degradation of the cell wall during the secretory cavity formation in Citrus grandis 'Tomentosa' fruits.

MAIN CONCLUSION: CgPG21 is mainly located in the cell wall, participates in the intercellular layer degradation of the cell wall during the formation of secretory cavity in the intercellular space-forming and lumen-expanding stages. The secretory cavity is a common structure in Citrus plants and is the main site for synthesis and accumulation of medicinal ingredients. The secretory cavity is formed in lysogenesis, when epithelial cells enter a process of programmed cell death. Pectinases are known to be involved in degradation of the cell wall during the cytolysis of secretory cavity cells, but the changes in cell structure, the dynamic characteristics of cell wall polysaccharides and the related genes regulating cell wall degradation are unclear. In this study, electron microscopy and cell wall polysaccharide-labeling techniques were used to study the main characteristics of cell wall degradation of the secreting cavity of Citrus grandis 'Tomentosa' fruits. At the same time, the full CDS length of the pectinase gene CgPG21 was cloned, encoding a protein composed of 480 amino acids. CgPG21 is mainly located in the cell wall, participates in the degradation of the intercellular layer of the cell wall during the development of the secretory cavity, and plays an important role in the formation of the secretory cavity in the intercellular space-forming and lumen-expanding stages. With the development of secretory cavity, the cell wall polysaccharides of epithelial cells gradually degrade. CgPG21 is mainly involved in the intercellular layer degradation.

Echinacea purpurea-derived homogeneous polysaccharide exerts anti-tumor efficacy via facilitating M1 macrophage polarization.

Echinacea purpurea modulates tumor progression, but the underlying mechanism is poorly defined. We isolated and purified a novel homogeneous polysaccharide from E. purpurea (EPPA), which was shown to be an arabinogalactan with a mean molecular mass (Mr) of 3.8 × 104 Da and with α- (1 → 5) -L-Arabinan as the backbone and α-L-Araf-(1→, →6)-ß-D-Galp-(1→, and →4)-α-D-GalpA-(1→ as the side chains. Interestingly, oral administration of EPPA suppresses tumor progression in vivo and shapes the immune cell profile (e.g., facilitating M1 macrophages) in tumor microenvironment by single-cell RNA sequencing (scRNA-seq) analysis. More importantly, EPPA activates the inflammasome through a phagocytosis-dependent mechanism and rewires transcriptomic and metabolic profile, thereby potentiating M1 macrophage polarization. Collectively, we propose that EPPA supplementation could function as an adjuvant therapeutic strategy for tumor suppression.

Lysine-40 succinylation of TAGLN2 induces glioma angiogenesis and tumor growth through regulating TMSB4X.

Protein lysine succinylation (Ksucc) represents an important regulatory mechanism of tumor development. In this work, the difference of protein Ksucc between HCMEC/D3 co-cultured with U87 (glioma endothelia cells, GEC) and without U87 (normal endothelia cells, NEC) was investigated using TMT labeling and affinity enrichment followed by high-resolution LC-MS/MS analysis. Interestingly, TAGLN2 was highly succinylated at K40 in GEC (15.36 folds vs. NEC). Compared to the Vector group, TAGLN2WT and a succinylation-mimetic TAGLN2K40E greatly promoted the angiogenesis of glioma in vitro and in vivo. Furthermore, the adhesion and metastasis of U87 co-cultured with GEC in the TAGLN2WT or TAGLN2K40E group were also significantly promoted. This was consistent with the increased expression of VE-cadherin and actin cytoskeleton remodeling induced by TAGLN2 K40succ in GEC. In addition, high K40succ of TAGLN2 was associated with poor prognosis in patients with glioma. Overexpression of TAGLN2K40E also markedly promoted the proliferation and migration of glioma cells, further analysis of in vivo xenograft tumors showed that there was a significant decrease in tumor size and angiogenesis in the TAGLN2K40R group. Notably, the co-localization of TMSB4X and TAGLN2 mainly in the nucleus and cytoplasm of glioma cells was detected by immunofluorescence staining. We identified TMSB4X as a potential target of TAGLN2, which was proved to interact with TAGLN2WT rather than TAGLN2K40A. And the inhibition of TMSB4X could markedly attenuate the proliferation and migration of glioma cells induced by TAGLN2 K40succ. The results revealed K40succ of TAGLN2 could be a novelty diagnosis and therapeutic target for gliomas.

Identification of novel indole derivatives as highly potent and efficacious LSD1 inhibitors.

Lysine-specific demethylase 1 (LSD1) is a FAD-dependent histone demethylase to catalyze the demethylation of H3K4 and H3K9 and thus is an attractive target for therapeutic cancer. Starting with a high micromolar compound 17i, structure-based optimization of novel indole derivatives is described by a bioelectronic isosteric strategy. Grounded by molecular modeling, medicinal chemistry has efficiently yielded low nanomolar LSD1 inhibitors. One of the compounds, B35, exhibited excellent LSD1 inhibition (IC50 = 0.050 ± 0.005 µM) and anti-proliferation against A549 cells (IC50 = 0.74 ± 0.14 µM). The further PK studies indicated compound B35 possessed favorable metabolic stability, in which the plasma t1/2 of p.o. and i.v. were 6.27 ± 0.72 h and 8.78 ± 1.31 h, respectively. Additionally, inhibitor B35 shows a strong antitumor effect and good safety in vivo. Meanwhile, compound B35 regulated genes are closely associated with transcriptional dislocation in cancer and PI3K/AKT pathway involving IGFBP3. Taken together, B35 could be a potent LSD1 inhibitor for further drug development.

Comparative analysis of absorbed ingredients and metabolites, and pharmacokinetic studies of Zhimu-Huangbai herb pair in the plasma of normal and type 2 diabetes mellitus rats by UHPLC-linear trap quadrupole-orbitrap MS and LC-MS/MS.

A new rapid ultra-high-performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry method was established for the qualitative analysis of absorbed ingredients and metabolites of Zhimu-Huangbai herb pair, which is used to treat type 2 diabetes mellitus. A total of 16 absorbed ingredients and 11 metabolites were identified in normal and type 2 diabetes mellitus rats, respectively. Such findings indicated that the diabetic model had no effect on the type of components in plasma. Seven absorbed ingredients and 11 metabolites were first identified after the oral administration of Zhimu-Huangbai herb pair. Thereafter, ultra-high-performance liquid chromatography coupled with linear trap quadrupole orbitrap mass spectrometry and liquid chromatography-API4000+ triple quadrupole mass spectrometer methods were established and validated for pharmacokinetic comparative studies of seven major bioactive components in normal and type 2 diabetes mellitus rats. Partial pharmacokinetic parameters in the plasma of type 2 diabetes mellitus rats were significantly different from those in normal rats. To our knowledge, this is the first comparison of absorbed ingredients and metabolites of Zhimu-Huangbai herb pair, and its use in pharmacokinetic studies between normal and type 2 diabetes mellitus rats. Ultimately, our findings provide insights into the clinical usage of Zhimu-Huangbai herb pair.

Design, synthesis and biological evaluation of novel benzofuran derivatives as potent LSD1 inhibitors.

Lysine-specific demethylase 1 (LSD1) is a FAD-dependent enzyme, which has been proposed as a promising target for therapeutic cancer. Herein, a series of benzofuran derivatives were designed, synthesized and biochemical evaluated as novel LSD1 inhibitors based on scaffold hopping and conformational restriction strategy. Most of the compounds potently suppressed the enzymatic activities of LSD1 and potently inhibited tumor cells proliferation. In particular, the representative compound 17i exhibited excellent LSD1 inhibition at the molecular levels with IC50 = 0.065 µM, as well as anti-proliferation against MCF-7, MGC-803, H460, A549 and THP-1 tumor cells with IC50 values of 2.90 ± 0.32, 5.85 ± 0.35, 2.06 ± 0.27, 5.74 ± 1.03 and 6.15 ± 0.49 µM, respectively. The binding modes of these compounds were rationalized by molecular docking. Meanwhile, a preliminary druggability evaluation showed that compound 17i displayed favorable liver microsomal stability and weak inhibitory activity against CYPs at 10 µM. Remarkably, H460 xenograft tumors studies revealed that 17i demonstrated robust in vivo antitumor efficacy without significant side effects. All the results demonstrated that compound 17i could represent a promising lead for further development.

QD-2: a novel open-framework aluminoborate with intersecting three-dimensional helical channels.

A novel open-framework aluminoborate, [CH3NH2(CH2)3NH ]-[AlB5O10] (QD-2), has been synthesized under hydrothermal conditions and characterized by IR, elemental analysis, thermogravimetric analysis, and powder and single-crystal X-ray diffractions. The compound crystallizes in the orthorhombic system, space group P2(1)2(1)2(1) (No. 19), a = 9.4352(4) A, b = 10.2972(2) A, c = 13.7511(5) A, V = 1336.0(8) A3, and Z = 4. Its structure consists of AlO4 tetrahedra and B5O10 clusters, which are interconnected to form the first example of a three-dimensional aluminoborate with intersecting helical channels.

Synthesis and characterization of QD-3: the first organically templated aluminoborate with 11-, 12-, and 14-ring intersecting channels.

QD-3, a novel aluminoborate (TETA) 2[Al 2B 10 O 20].0.25H 2O (TETA = triethylenetetramine), has been solvothermally synthesized and characterized by IR, thermogravimetric analysis, power X-ray diffraction, elemental analysis, and single-crystal X-ray diffraction. This compound crystallizes in the orthorhombic system, space group Pna2 1 (No. 33), a = 25.9174(2) A, b = 7.4028(5) A, c = 17.013(4) A, V = 3264.2(2) A (3), and Z = 4. Its structure consists of AlO 4 tetrahedra and B 5O 10 clusters, which are interconnected and form a three-dimensional framework with large 11-, 12-, and 14-ring channels.

Antioxidant capacity and lipophilic content of seaweeds collected from the Qingdao coastline.

Lipophilic extracts from 16 species of seaweeds collected along the Qingdao coastline were screened and evaluated for their antioxidant activities (AA) using the beta-carotene-linoleate assay system. The diethyl ether soluble extracts of all selected seaweeds exhibited various degrees of antioxidative efficacy in each screen. The highest antioxidant capacities among the tested samples were observed for Rhodomela confervoides and Symphyocladia latiuscula and were comparable with that of the well-known antioxidant butylated hydroxytoluene and greater than that of propyl gallate. The lipophilic content of all 16 samples and the chemical composition of 4 selected seaweeds, R. confervoidesand S. latiuscula, which had higher AA, Laminaria japonica, which had intermediate AA, and Plocamium telfairiae, which had lower AA, were analyzed by gas chromatography and gas chromatography-mass spectrometry, respectively. Fatty acids and alkanes were found. The present data indicated an increase in antioxidative property with increasing content of unsaturated fatty acid. The result of this study suggests that seaweeds can be considered as a potential source for the extraction of lipophilic antioxidants, which might be used as dietary supplements or in production in the food industry. This is the first report on the antioxidant activities of lipophilic extracts from seaweeds.