Total Synthesis of Ganoderma Meroterpenoids Cochlearol B and Its Congeners Driven by Structural Similarity and Biological Homology.

Secondary metabolites that have the same biological origin must share some relationship in their biosynthesis. Exploring this relationship has always been a significant task for synthetic biologists. However, from the perspective of synthetic chemists, it is equally important to propose, prove, or refute potential biosynthetic pathways in order to elucidate and understand the biosynthesis of homologous secondary metabolites. In this study, driven by the high structural similarity between the homologous Ganoderma meroterpenoids cochlearol B and ganocin B, two chemically synthetic strategies were designed and investigated sequentially for the synthesis of cochlearol B from ganocin B. These strategies include intramolecular metal-catalyzed hydrogen atom transfer (MHAT) and intramolecular photochemical [2+2] cycloaddition. The aim was to reveal their potential biosynthetic conversion relationship using chemical synthesis methods. As a result, a highly efficient total synthesis of cochlearol B, cochlearol T, cochlearol F, as well as the formal total synthesis of ganocins A-B, and ganocochlearins C-D, has been achieved. Additionally, a novel synthetic approach for the synthesis of 6,6-disubstituted 6H-dibenzo[b,d]pyran and its analogues has been developed through palladium(II)-catalyzed Wacker-type/cross-coupling cascade reactions.

Total Synthesis of Ryanodane Diterpenoids Garajonone and 3-epi-Garajonone.

Ryanodane diterpenes are structurally complex natural products that are well-known for their high degree of oxidation and the challenges associated with synthesizing them within the terpene class. Herein, we present a two-stage synthetic strategy that draws inspiration from the broad biosynthesis of terpenes, allowing us to successfully achieve the first chemical synthesis of garajonone, a ryanodane diterpenoid that occurs naturally at low abundance, as well as its epimer, 3-epi-garajonone. The key to this success lies in the rapid construction of the carbon framework of target molecule by employing an early-stage palladium-catalyzed Heck/carbonylative esterification cascade annulation, followed by successive late-stage selective redox manipulation to establish the desired oxidation state of the molecule. This research not only showcases the synthesis of garajonone and its epimer but also provides a platform for the chemical synthesis of other members and analogs within this complex diterpenoid family.

Bioinspired Total Synthesis of Cephalotaxus Diterpenoids and Their Structural Analogues.

Herein, we present a unified chemical synthesis of three subgroups of cephalotaxus diterpenoids. Key to the success lies in adopting a synthetic strategy that is inspired by biosynthesis but is opposite in nature. By employing selective one-carbon introduction and ring expansion operations, we have successfully converted cephalotane-type C18 dinorditerpenoids (using cephanolide B as a starting material) into troponoid-type C19 norditerpenoids and intact cephalotane-type C20 diterpenoids. This synthetic approach has enabled us to synthesize cephinoid H, 13-oxo-cephinoid H, 7-oxo-cephinoid H, fortalpinoid C, 7-epi-fortalpinoid C, cephanolide E, and 13-epi-cephanolide E. Furthermore, through the development of an intermolecular asymmetric Michael reaction between ß-oxo esters and ß-substituted enones, we have achieved the enantioselective synthesis of advanced intermediates within our synthetic sequence, thus formally realizing the asymmetric total synthesis of the cephalotaxus diterpenoids family.

Reduced Volume Expansion of Micron-Sized SiOx via Closed-Nanopore Structure Constructed by Mg-Induced Elemental Segregation.

The inherently huge volume expansion during Li uptake has hindered the use of Si-based anodes in high-energy lithium-ion batteries. While some pore-forming and nano-architecting strategies show promises to effectively buffer the volume change, other parameters essential for practical electrode fabrication, such as compaction density, are often compromised. Here we propose a new in situ Mg doping strategy to form closed-nanopore structure into a micron-sized SiOx particle at a high bulk density. The doped Mg atoms promote the segregation of O, so that high-density magnesium silicates form to generate closed nanopores. By altering the mass content of Mg dopant, the average radii (ranged from 5.4 to 9.7 nm) and porosities (ranged from 1.4 % to 15.9 %) of the closed pores are precisely adjustable, which accounts for volume expansion of SiOx from 77.8 % to 22.2 % at the minimum. Benefited from the small volume variation, the Mg-doped micron-SiOx anode demonstrates improved Li storage performance towards realization of a 700-(dis)charge-cycle, 11-Ah-pouch-type cell at a capacity retention of >80 %. This work offers insights into reasonable design of the internal structure of micron-sized SiOx and other materials that undergo conversion or alloying reactions with drastic volume change, to enable high-energy batteries with stable electrochemistry.

Total Synthesis of Metaphanine and Oxoepistephamiersine.

Herein, we report a concise and divergent synthesis of the complex hasubanan alkaloids metaphanine and oxoepistephamiersine from commercially available and inexpensive cyclohexanedione monoethylene acetal. Our synthesis features a palladium-catalyzed cascade cyclization reaction to set the tricyclic carbon framework of the desired molecules, a regioselective Baeyer-Villiger oxidation followed by a MeNH2 triggered skeletal reorganization cascade to construct the benzannulated aza[4.4.3]propellane, and a strategically late-stage regio-/diastereoselective oxidative annulation of sp3 C-H bond to form the challenging THF ring system and hemiketal moiety in a single step. In addition, a highly enantioselective alkylation of cyclohexanedione monoethylene acetal paved the way for the asymmetric synthesis of target molecular.

Insights into Anion-Solvent Interactions to Boost Stable Operation of Ether-Based Electrolytes in Pure-SiOx ||LiNi0.8 Mn0.1 Co0.1 O2 Full Cells.

Ether solvents with superior reductive stability promise excellent interphasial stability with high-capacity anodes while the limited oxidative resistance hinders their high-voltage operation. Extending the intrinsic electrochemical stability of ether-based electrolytes to construct stable-cycling high-energy-density lithium-ion batteries is challenging but rewarding. Herein, the anion-solvent interactions were concerned as the key point to optimize the anodic stability of the ether-based electrolytes and an optimized interphase was realized on both pure-SiOx anodes and LiNi0.8 Mn0.1 Co0.1 O2 cathodes. Specifically, the small-anion-size LiNO3 and tetrahydrofuran with high dipole moment to dielectric constant ratio realized strengthened anion-solvent interactions, which enhance the oxidative stability of the electrolyte. The designed ether-based electrolyte enabled a stable cycling performance over 500 cycles in pure-SiOx ||LiNi0.8 Mn0.1 Co0.1 O2 full cell, demonstrating its superior practical prospects. This work provides new insight into the design of new electrolytes for emerging high-energy density lithium-ion batteries through the regulation of interactions between species in electrolytes.

Hsa-miRNA-143-3p regulates the odontogenic differentiation of human stem cells from the apical papilla by targeting NFIC.

AIM: To evaluate the effects of hsa-miRNA-143-3p on the cytodifferentiation of human stem cells from the apical papilla (hSCAPs) and the post-transcriptional regulation of Nuclear factor I-C (NFIC). METHODOLOGY: miRNA expression profiles in human immature permanent teeth and during hSCAP differentiation were examined. hSCAPs were treated with miR-143-3p overexpression or silencing viruses, and the proliferation and odontogenic and osteogenic differentiation of these stem cells, and the involvement of the NFIC pathway, were investigated. Luciferase reporter and NFIC mutant plasmids were used to confirm NFIC mRNA as a direct target of miR-143-3p. NFIC expression analysis in the miR-143-3p overexpressing hSCAPs was used to investigate whether miR-143-3p functioned by targeting NFIC. Student's t-test and chi-square tests were used for statistical analysis. RESULTS: miR-143-3p expression was screened by microarray profiling and was found to be significantly reduced during hSCAP differentiation (p < .05). Overexpression of miR-143-3p inhibited the mineralization of hSCAPs significantly (p < .05) and downregulated the levels of odontogenic differentiation markers (NFIC [p < .05], DSP [p < .01] and KLF4 [p < .01]), whereas silencing of miR-143-3p had the opposite effect. The luciferase reporter gene detection and bioinformatic approaches identified NFIC mRNA as a potential target of miR-143-3p. NFIC overexpression reversed the inhibitory effect of miR-143-3p on the odontogenic differentiation of hSCAPs. CONCLUSIONS: miR-143-3p maintained the stemness of hSCAPs and modulated their differentiation negatively by directly targeting NFIC. Thus, inhibition of this miRNA represents a potential strategy to promote the regeneration of damaged tooth roots.

Total Synthesis of Sieboldine A.

Previously, we have finished the total synthesis of lycojaponicumin A (2) via development of an efficient synthetic strategy using semipinacol rearrangement as a key step. In order to further demonstrate the generality of this synthetic route, herein, we report the total synthesis of another fawcettimine-type alkaloid sieboldine A (1) from the same intermediate, which possesses an A/B/D tricyclic ring system and vicinal quaternary centers of 1. The synthesis features late-stage site-selective redox reactions, Schmidt glycosylation cyclization, and highly selective transformations.

Divergent Biomimetic Total Syntheses of Ganocins†A-C, Ganocochlearins†C and D, and Cochlearol†T.

A divergent synthetic approach to six Ganoderma meroterpenoids, namely ganocins A-C, ganocochlearins C and D, and cochlearol T, has been developed for the first time. This synthetic route features a two-phase strategy which includes early-stage rapid construction of a common planar tricyclic intermediate followed by highly selective late-stage transformations into various Ganoderma meroterpenoids. Key to the strategy are a bioinspired intramolecular hetero-Diels-Alder reaction and Stahl-type oxidative aromatization, allowing efficient formation of the common tricyclic phenol intermediate. A nucleophilic dearomatization of the phenol unit, combined with a regioselective 1,4-reduction of the resulting dienone, enabled rapid access to ganocins B and C. Additionally, site-selective Mukaiyama hydration, followed by an intramolecular oxa-Michael addition/triflation cascade, served as a key strategic element in the chemical synthesis of ganocin A.

Total Synthesis of (±)-Cephanolides B and C via a Palladium-Catalyzed Cascade Cyclization and Late-Stage sp3 C-H Bond Oxidation.

Herein, we report the first total syntheses of complex cephalotaxus diterpenoids cephanolide B and C from commercially available 5-bromo-2-methylanisole. Key to the success of this synthetic route is a palladium-catalyzed cascade cyclization reaction, which allowed us to efficiently forge the 6-5-6 cis-fused tricyclic ring systems found in the entire family of cephalotaxus diterpenoids. Additionally, site-selective late-stage sp3 C-H bond oxidation served as a key strategic element in the chemical synthesis of cephanolide C.

Bifunctional Furfuryl Cations Strategy: Three-Component Synthesis of Enamidyl Triazoles.

A new multicomponent synthesis of functionalized enamidyl triazoles starting from simple and readily available starting materials is described. A simple treatment of a dichloromethane solution of an azide, amine, and 5-bromo-2-furylcarbinol with a Lewis acid provides the enamidyl triazole in good to high yield. A triple domino sequence, formal [3+2] cycloaddition/ring-opening/amidation, is involved in this new skeleton-generating reaction.

Neuroprotective effect of a novel Chinese herbal decoction on cultured neurons and cerebral ischemic rats.

BACKGROUND: Historically, traditional Chinese medicine has been widely used to treat stroke. Based on the theory of Chinese medicine and the modern pharmacological knowledge of herbal medicines, we have designed a neuroprotective formula called Post-Stroke Rehabilitation (PSR), comprising seven herbs - Astragalus membranaceus (Fisch.) Bunge, Salvia miltiorrhiza Bunge, Paeonia lactiflora Pall., Cassia obtusifolia L., Ligusticum chuanxiong Hort., Angelica sinensis (Oliv.) Diels, and Glycyrrhiza uralensis Fisch. We aim to examine the neuroprotective activity of PSR in vitro and in vivo, and to explore the underlying molecular mechanisms, to better understand its therapeutic effect and to further optimize its efficacy. METHODS: PSR extract or vehicle was applied to primary rat neurons to examine their survival effects against N-methyl-D-aspartate (NMDA)-elicited excitotoxicity. Whole-cell patch-clamp recording was conducted to examine the NMDA-induced current in the presence of PSR. ERK- and CREB-activation were revealed by western blot analysis. Furthermore, PSR was tested for CRE promoter activation in neurons transfected with a luciferase reporter. The protective effect of PSR was then studied in the rat middle cerebral artery occlusion (MCAO) model. MCAO rats were either treated with PSR extract or vehicle, and their neurobehavioral deficit and cerebral infarct were evaluated. Statistical differences were analyzed by ANOVA or t-test. RESULTS: PSR prominently reduced the death of cultured neurons caused by NMDA excitotoxicity in a dose-dependent manner, indicating its neuroprotective property. Furthermore, PSR significantly reduced NMDA-evoked current reversibly and activated phosphorylation of ERK and CREB with distinct time courses, with the latter's kinetics slower. PSR also triggered CRE-promoter activity as revealed by the increased expression of luciferase reporter in transfected neurons. PSR effectively reduced cerebral infarct and deficit in neurological behavior in MCAO rats when PSR decoction was administered starting either 6 days before or 6 h after onset of ischemia. CONCLUSIONS: PSR is neuroprotective both in vitro and in vivo - it protects cultured neurons against NMDA excitotoxicity, and effectively reduces ischemic injury and neurobehavioral deficit in MCAO rats in both the pre- and post-treatment regimens. The underlying neuroprotective mechanisms may involve inhibition of NMDA receptor current and activation of ERK and CREB. This study provides important preclinical data necessary for the further development of PSR for stroke treatment.

[Effects of stromal cell-derived factor-1 on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cells].

OBJECTIVE: To compare the effects of stromal cell-derived factor-1 (SDF-1) and granulocyte colony-stimulating factor (G-CSF) on proliferation, migration, and odontoblastic differentiation of human dental pulp stem cell (DPSC) in vitro. METHODS: DPSCs were cultured in vitro and treated with either 100 µg/L SDF-1 or 100 µg/L G-CSF. Cell counting kit-8 (CCK-8) and colony-forming unit (CFU) were used to detect the effect of SDF-1 and G -CSF on the proliferation ability of DPSC. Cell migration of DPSC was determined by wound healing assay and Transwell migration assay. The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP) staining, ALP activity and alizarin red S staining. The expression of odontoblastic-related genes such as dentin matrix protein 1 (DMP-1) and dentin sialophosphoprotein (DSPP) were quantified by real-time RT-PCR. RESULTS: SDF-1 and G-CSF promoted the proliferation of DPSC slightly, but the difference was not statistically significant. Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01), but there was no significant difference between the two factors. In Transwell migration assay, the number of migrated cells of the control group was 5.0 ± 1.4 per sight, while the SDF-1 group was 24.3 ± 6.8 per sight and the G-CSF group was 11.8 ± 3.3 per sight, suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF, and SDF-1 was more effective than G-CSF (P<0.05). Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining. Higher ALP activity, more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment. CONCLUSION: SDF-1 had no significant effect on the proliferation of DPSC, but could significantly promote cell migration and odontoblastic differentiation of DPSC. Its effect on DPSC was better than G-CSF.

[A retrospective study on pulpal tissue prognosis of avulsed permanent teeth in children].

OBJECTIVE: To analyze the pulpal prognosis of replanted permanent teeth in children and to examine the associated factors. METHODS: The samples consisted of 67 children with 81 avulsed permanent teeth treated in Peking University Hospital of Stomatology from 2000 to 2012. Their clinical data of the replanted teeth were collected, and the follow-up period was no less than 12 months. The pulpal prognosis was classified as pulp necrosis and pulp non-necrosis. RESULTS: The replantation within 30 minutes after avulsion led to the most significant increase in pulpal healing (P<0.05). The dental pulp of 80% (4/5) young permanent teeth replanted within 30 minutes remained vital, while all the teeth replanted after 30 minutes developed pulp necrosis within 60 days after replantation. CONCLUSION: The extra-alveolar period significantly affects the prognosis of pulp in immature permanent teeth after replantation. When the extra-alveolar period is more than 30 minutes, the chance of pulp revascularization after replantation is very low, therefore pulp extirpation should be performed.

[Spatiotemporal expression pattern of E-cadherin and P-cadherin during mouse tooth development].

OBJECTIVE: To investigate the expression patterns of E-cadherin and P-cadherin in murine-tooth germs at early developmental stages. METHODS: Mandible samples of CD1 mice from embryonic day 12.5 to postnatal day 3.5 were collected. The expressions of E-cadherin and P-cadherin in murine mandibular first molar germs were detected by immunofluorescence and observed under confocal fluorescence microscope. HE staining was performed for tissue morphology. RESULTS: Both E-cadherin and P-cadherin were widely expressed in the epithelial tissues through early developmental stages. The E-cadherin expression was increased in polarizing pre-ameloblasts, whereas the P-cadherin expression declined. The expression of the P-cadherin could be detected in epithelial tissues before bud stage, and expressed in mature ameloblasts at secretory stage. CONCLUSION: The E-cadherin and P-cadherin expressed in different spatiotemporal expression patterns, indicating their individual functions during tooth development. P-cadherin might function in the secretion and mineralization of enamel.

Short, enantioselective total synthesis of chatancin.

An enantioselective total synthesis of the polycyclic diterpene (+)-chatancin, a potent PAF antagonist, is reported. Proceeding in seven steps from dihydrofarnesal, this synthetic route was designed to circumvent macrocyclization-based strategies to complex, cyclized cembranoids. The described synthesis requires only six chromatographic purifications, is high yielding, and avoids protecting-group manipulations. An X-ray crystal structure of this fragile marine natural product was obtained.

Nuclear factor I-C expression pattern in developing teeth and its important role in odontogenic differentiation of human molar stem cells from the apical papilla.

Nuclear factor I-C (NFIC) has an important role in the development of murine dental roots, but its role in human root formation is unreported. We thus elucidated the regulatory role of NFIC in the differentiation of human stem cells from the apical papilla (hSCAPs). The first step for this was to determine the expression of NFIC in human teeth, and it was found that NFIC expression was restricted to the odontoblasts and preodontoblasts of the developing molars of humans and mice. NFIC was found to be expressed in odontoblast-like cells after the subcutaneous transplantation of hSCAPs. NFIC expression was concomitant with dentin sialophosphoprotein (DSPP) in the mineralization of hSCAPs. NFIC knockdown in hSCAPs significantly inhibited expression of DSPP and promoted that of dentin matrix protein 1 (DMP1), meanwhile upregulated the expression of TGF-ß1 and downregulated SMAD3 and SMAD4. NFIC expression was significantly upregulated after TGF-ß1 treatment in hSCAPs. NFIC knockdown prolonged G1 phase of the cell cycle, but had no effect on cell proliferation and migration. These results suggest that NFIC is involved in the development of human root dentin and the regulation of odontoblastic differentiation of hSCAPs. NFIC may participate in the DMP1-DSPP signaling pathway and comprises a complex signaling cycle with TGF-ß1.

Bioinspired Collective Total Synthesis of (±)-Rhynchines A-E.

Herein, we present the first racemic total synthesis of the structurally complex monoterpene indole alkaloids rhynchines A-E, starting from commercially available methyl nicotinate and 3-(2-bromoethyl)-1H-indole. The success of our synthesis is attributed to the utilization of a bioinspired synthetic strategy, which facilitated the rapid construction of the pentacyclic core skeleton of the target molecules through biomimetic skeletal rearrangement and late-stage C-H oxidative cyclization. Additionally, silica-gel-promoted tautomerization played a crucial role as a strategic element in the chemical synthesis of rhynchines A and B.

Msx1 regulates proliferation and differentiation of mouse dental mesenchymal cells in culture.

The homeobox, msh-like 1 (MSX1) protein is essential for cell proliferation and differentiation. Tooth germ development of Msx1 knockout mouse is arrested at the bud stage, impeding an understanding of its role beyond this stage of tooth development. The aims of this study were to investigate the potential role of MSX1 in the regulation of proliferation and differentiation of dental mesenchymal cells in culture, and to preliminarily explore its underlying mechanism of action. Tooth germs were isolated from embryonic day (E)15.5 mice. The mesenchyme was separated and digested into a single-cell suspension, and then cultured in vitro. Isolated dental mesenchymal cells were transfected with MSX1 small interfering RNA, and the effects on cell proliferation, cell cycle distribution, and the expression of bone morphogenetic protein 2 (Bmp2) and bone morphogenetic protein 4 (Bmp4) were studied. We also compared the expression levels of alkaline phosphatase (Alp), type I collagen (Col1A), osteocalcin (Ocn), runt-related transcription factor 2 (Runx2), dentin sialophosphoprotein (Dspp) and dentin matrix protein 1 (Dmp1), and mineralized nodule formation, between control and MSX1 siRNA-transfected groups after the induction of odontoblast differentiation. Knockdown of Msx1 expression was associated with decreased cell proliferation, prolonged time in the S phase of the cell cycle, enhanced odontoblast differentiation, and elevated Bmp2 and Bmp4 expression. We conclude that MSX1 may promote proliferation and prevent the differentiation of dental mesenchymal cells by the inhibition of Bmp2 and Bmp4 expression.

Effect of skimmed pasteurized milk and Hank's balanced salt solution on viability and osteogenic differentiation of human periodontal ligament stem cells.

BACKGROUND/AIM: The purpose of this study was to compare the effect of skimmed pasteurized milk and Hank's balanced salt solution on the viability and osteogenic differentiation potential of the human periodontal ligament stem cells at room temperature in vitro. MATERIAL AND METHODS: Human periodontal ligament stem cells were obtained from extracted healthy third molars and conserved in skimmed pasteurized milk and Hank's balanced salt solution for 1, 2, and 4 h at room temperature to detect the viability of the cells and their osteogenic differentiation potential. RESULTS: The efficacy of skimmed pasteurized milk on cell viability at 4 h was significantly higher than that of HBSS (P < 0.05), and cells stored in skimmed pasteurized milk showed significantly higher levels of mineralization than those in HBSS at 2 and 4 h (P < 0.05). CONCLUSIONS: Skimmed pasteurized milk was more effective than Hank's balanced salt solution in maintaining the viability and osteogenic differentiation potential of PDLSCs at room temperature in vitro.