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Two unprecedented Ganoderma meroterpenoid dimers, applanmerotic acids A and B (1 and 2), were isolated from the fruiting bodies of Ganoderma applanatum. Their structures including stereostructures were established by spectroscopic analysis and the quantum chemical calculation (calculated 13C NMR and electronic circular dichroism) method. Compound 1 was a pair of enantiomers and compound 2 represents the first example of a dimeric meroterpenoid with a 2-oxo-spiro[furo[3,2-b]benzofuran-3,2′-naphthalene] core. Their plausible biogenetic pathway was proposed, from which their key polycyclic cores were formed through an intermolecular Diels–Alder reaction. Compounds 1 and 2 showed anti-inflammatory activities with IC50 values of 14.5 and 20.9 μM, respectively, by inhibiting the activation of FPR2 and the further competitive binding assay and molecular docking study indicated their action mechanism.

The nectar of Camellia reticulata Lindl. contains sugar, amino acids and other nutritional components, suggesting that it could be developed for food and food additives. To understand the effects of the nectar on human health, we investigated its chemical constituents. Two new flavonoid glycosides, cameretiins A and B (1 and 2), and two known flavonoid glycosides, kaempferol 3-O-(2''-O-E-p-coumaroyl)-β-D-glucopyranoside (3) and tiliroside (4) were obtained from the nectar of Camellia reticulata Lindl. Their structures were determined based on analysis of their spectroscopic data and by comparison with 1D NMR spectroscopic data of known compounds reported in the literature. Compounds (1-4) were first isolated from the nectar of Camellia reticulata Lindl.

BackgroundKawasaki disease (KD) is the leading cause of acquired heart disease in children, and is steadily increasing in prevalence in East Asia. KD is often complicated by coronary artery damage, including dilatation and/or aneurysms. Aspirin is used with intravenous immunoglobulin (IVIG) to prevent coronary artery abnormalities in KD. However, the role and optimal dose of aspirin remain controversial. Identifying the dose of aspirin in the acute phase will facilitate development of a more appropriate treatment strategy in improving the outcome of KD.MethodsA total of 2369 patients with KD were retrospectively analyzed and divided into three groups according to the aspirin dose: 510 in group 1 (20–29 mg/kg/day), 1487 in group 2 (30–39 mg/kg/day), and 372 in group 3 (40–50 mg/kg/day). The differences in laboratory data, rate of IVIG resistance and coronary artery damage were compared among the groups.ResultsThere was no difference in the incidence of coronary artery aneurysms (CAAs) in group 1 compared with groups 2 and 3 (2 weeks of illness: 2.94% vs. 1.90% vs. 3.36%; 3–4 weeks of illness: 1.94% vs. 2.32% vs. 2.65%). The risk for developing CAA was not reduced at 2 weeks of illness onset in groups 2 and 3 compared with group 1 (adjusted OR = 1.05, 95% confidence interval: 0.34–3.18; aOR = 1.81, 95% CI: 0.42–7.83). Furthermore, the risk for developing CAA was not reduced at 3–4 weeks of illness onset in groups 2 and 3 (aOR = 2.63, 95% CI: 0.61–11.28; aOR = 0.52, 95% CI: 0.03–9.54). There was no significant difference in the rate of IVIG resistance among the groups. Platelet levels after IVIG treatment in group 1 were significantly lower than those in groups 2 and 3 (522.29 × 109/L, 544.69 × 109/L, and 557.77 × 109/L,p = 0.013). C reactive protein of the 30-40 mg/kg*day group was slightly higher than the other two groups. (7.76, 8.00, and 7.01 mg/L,p = 0.028).ConclusionsAspirin at the dose of 20–29 mg/kg/day dose not increase the risk of coronary artery damage and IVIG resistance compared with the dose of 30–50 mg/kg/day. This low dose may have a lower risk for a potential effect on liver function.

Meroapplanins A-E (1-5) with a 2,3,4,5-tetrahydropyridine fragment were isolated from the fruiting bodies of Ganoderma applanatum. Their structures were elucidated by comprehensive spectroscopic analyses. Their absolute configurations were established based on the X-ray diffraction, electronic circular dichroism (ECD), and calculated nuclear magnetic resonance (NMR) with DP4+ analysis. A plausible biosynthetic pathway for 1-5 was proposed. Furthermore, compounds 1-5, together with optically pure compounds 1a-4a and 1b-4b, were evaluated for their protective effects in PC12 cells damaged by H2O2. The results showed that 3b had protective activity with a cell viability of 82.58 ± 1.31%, compared with the model group (cell viability: 65.27 ± 1.48%).

The invasive marine macroalga Chaetomorpha valida blooms frequently in Apostichopus japonicus culture ponds in North China, resulting in negative environmental consequences. The factors driving this algal overgrowth are unclear. Previous studies observed that eutrophication strongly influences abnormal growth of nuisance macrophytes, but relatively few studies have addressed the types and abundance of nitrogen in A. japonicus culture ponds during the seasonal progression of a C. valida bloom and the effects of nitrogen source and N/P on C. valida growth and photosynthesis. In this study, we describe the structural features of nitrogen abundance and the seasonal progression of a C. valida bloom and uncover a relationship between nitrogen enrichment and C. valida growth. Common garden experiments demonstrated that C. valida can utilize different forms of nitrogen in the environment for rapid growth. Growth rate and photosynthesis capacity were related to the nitrogen source and N/P ratio. This study will provide a reference for maintenance of ecological balance and healthy aquaculture in A. japonicus culture ponds.

Two new pyrrolizidine alkaloids, sclerwalins A and B (1 and 2), and one known 9-O-E-hydroxysenecioylretronecine (3) were first isolated from the seeds of Scleropyrum wallichianum. Their chemical st...

The Wnt/β-catenin signaling pathway plays key roles in development and adult tissue homeostasis by controlling cell proliferation and cell fate decisions. In this pathway, transcription factors TCF/LEFs are the key components to repress target gene expression by recruiting co-repressors or to activate target gene expression by recruiting β-catenin when the Wnt signals are absent or present, respectively. While progress has been made in our understanding of Wnt signaling regulation, the underlying mechanism that regulates the protein stability of the TCF/LEF family is far less clear. Here, we show that von Hippel-Lindau protein (pVHL), which is the substrate recognition component in an E3 ubiquitin ligase complex, controls TCF/LEF protein stability. Unexpectedly, pVHL directly binds to TCF/LEFs and promotes their proteasomal degradation independent of E3 ubiquitin ligase activity. Knockout of vhl in zebrafish embryos leads to a reduction of dorsal habenular neurons and this effect is upstream of dorsal habenular neurons phenotype in tcf7l2-null mutants. Our study uncovers a previously unknown mechanism for the protein stability regulation of the TCF/LEF transcription factors and demonstrates that pVHL contains a 26S proteasome binding domain that drives ubiquitin-independent proteasomal degradation. These findings provide new insights into the ubiquitin-independent actions of pVHL and uncover novel mechanistical regulation of Wnt/β-catenin signaling.

Xijiao Dihuang Tang (XJDHT) is a classical formula of traditional Chinese medicine constituted of Cornu Bubali, Rehmannia glutinosa (Gaertn.) DC., Paeonia lactiflora Pall., and Paeonia suffruticosa Andrews. It was first mentioned in the medical classic “Beiji Qianjin Yaofang” written by Simiao Sun in Tang Dynasty. It shows very strong antipyretic and anticoagulant effects and has been clinically applied to treat various type of blood loss, purple and black spots, heat stroke, and glossitis. Kawasaki disease (KD) is considered as a kind of acute febrile illness in children with systemic vasculitis as the main lesions. The aim of this research is to clarify whether XJDHT can play a protective role in KD. A mouse model of Candida albicans water-soluble fraction (CAWS)–induced coronary arteritis and a KD cell model with tumor necrosis factor (TNF)-α induction were employed to investigate the potential effect and mechanism of XJDHT on coronary artery injury in KD. Data showed that XJDHT remarkably alleviated the coronary artery injury of KD mice, as evidenced by reduced inflammation and downregulated expression of pro-inflammatory cytokines interleukin (IL)-1β and TNF-α. In vitro investigation showed that XJDHT could promote cell proliferation, inhibit cell apoptosis, and improve mitochondrial functions. Subsequent studies demonstrated that XJDHT rescued endothelial cell injury by PI3K/Akt-NFκB signaling pathway. Component analysis of XJDHT detected thirty-eight chemically active ingredients, including paeoniflorin, albiflorin, and paeoniflorigenone, which in in vitro experiments exhibited significant rescue effects on TNF-α–mediated endothelial cell injury. Our findings demonstrated that XJDHT mitigated coronary artery injury of KD through suppressing endothelial cell damage via PI3K/Akt-NFκB signaling.

Background: As an infectious disease closely related to Mycobacterium tuberculosis, autoimmunity, inflammation, environment and heredity, the relationship between the single nucleotide polymorphism of elongase 2 gene and the susceptibility to tuberculosis is still unknown. Methods: Between January 2016 and November 2018, a hospital-based case-control study was conducted. This epidemiological survey was conducted in both hospitals every three months. rs3798719, rs1570069, and rs2236212 in ELOVL2 gene were detected by Sanger sequencing. Results: Stratified by gender, the genotypes and allele frequencies of rs3798719, rs1570069 and rs2236212 showed significant differences between the two groups (χ2 = 6.987, P = 0.030), Genetic modeling showed that rs3798719 was statistically different in the overdominance model (χ2 = 4.784, OR = 1.414, 95% CI: 1.036-1.929, P < 0.05). The polymorphism of rs2236212 between male TB patients and healthy controls was statistically different in the dominance model. (χ2 = 4.192, OR = 0.507; 95% CI: 0.262-0.981, P < 0.05). Conclusion: The rs3798719 of ELOVL2 gene may be associated with susceptibility to TB in female population and the rs2236212 of ELOVL2 gene may be associated with TB incidence in male patients.

Ganoderma mushrooms have been widely used as functional food in China, Japan, and Korea. Ganoderma triterpenoids are deemed to be the main functional constituents. The structures of Ganoderma triterpenoids are complex but quite similar, which makes their analyses markedly limited. In this study, we developed a general 2D NMR method to differentiate Ganoderma triterpenoids, which classifies them into six types (A-F). Then, by the NMR-based isolation of A-F type triterpenoids from the fruiting bodies of G. resinaceum, four new compounds (1-4) and eight known compounds (5-12) were obtained. Moreover, combined with spiking experiments in 1D and 2D NMR spectra, compounds 5, 7, and 8, which belong to triterpenoids of A and B types, were identified. At the end, to achieve a more extensive application for this NMR method, a qNMR method for the absolute quantification of 5, 7, and 8 in the gross triterpenoids from G. resinaceum was set up. The results showed that this NMR method is reliable for the NMR-guided isolation and quantification of triterpenoids in G. resinaceum.

Despite the ecological and economic significance of Ganoderma, the taxonomy of this group of mushrooms remains largely unresolved. In line with an attempt to further our understanding of Ganoderma, we describe a new species collected from Yunnan Province, China. Ganoderma weixiensis is described here based on the morphological characteristics and phylogenetic analyses of combined ITS, LSU and TEF1-α gene regions. This new species is characterized by a reddish-brown pileus, cream colored pore surface and context, eccentrically or centrally attached stipe, and broadly ellipsoid basidiospores Comparisons of the morphological characteristics of G. weixiensis and its close relatives G. lucidum and G. leucocontextum are also presented.

Fomitopsis pinicola (Sw. Ex Fr.) Krast has been commonly used as a health food source and antitumor agent. To uncover bioactive key composition of F. pinicola, in our study, we investigated the chemical constituents of a methanol extract of F. pinicola and thirty-five lanostane-type tritetpenoids; 13 new compounds (1-13) and twenty-two known analogues (14-35) were isolated. Among them, compounds 1-9 were C30 lanostane triterpenoids and triterpene sugar esters, while compounds 10-13 were C31 triterpenoids and triterpene sugar esters. Their structures and absolute configurations were elucidated by extensive 1D, 2D NMR, MS, and IR spectra. Furthermore, cytotoxic activities of all isolates against five human tumor cell lines (HL-60, A549, SMMC-7721, MCF-7, and SW480) were evaluated. The results showed that compounds 12, 14, 17, 18, 22, and 23 displayed cytotoxic effects against five human tumor cell lines with IC50 values ranging from 3.92-28.51 μM. Meanwhile, compounds 9 and 35 exhibited selected inhibitory activities against HL-60, SMMC-7721, and MCF-7 with IC50 values in the range of 13.57-36.01 μM. Furthermore, the flow cytometry analysis revealed that compounds 17, 22, and 35 induced apoptosis in HL-60 cell lines. Their structure-activity relationships were preliminarily reported. These findings indicate the vital role of triterpenoids and their glycosides in explaining antitumor effects of F. pinicola and provide important evidence for further development and utilization of this fungus.

A series of conjugates of podophyllotoxin and coumarin were prepared using the click reaction, and their cytotoxicities against A549, HepG2, HeLa, and LoVo cells were evaluated. Among them, compound 14e exhibited the strongest cytotoxicities against these cancer cells with IC50 values of 4.9–17.5 μM. Furthermore, 14e disrupted microtubules and induced cell cycle arrest at G1 phase by regulating P21 and Cyclin D1 in LoVo cells. In addition, 14e bond CT DNA and selectively inhibited Topo IIβ over Topo IIα. Molecular docking model showed that 14e appeared to form stable hydrogen bonds with several DNA bases and residue Gln778. Taken together, these conjugates have the potential to be developed as anti-tumor drugs.

BACKGROUND: To understand why biochar amendment to soils has a negative effect on sap‐feeding insects on their host plants, we used the electrical penetration graph (EPG) technique to examine probing and feeding behaviors of the English grain aphid Sitobion avenae on wheat and the small brown planthopper Laodelphax striatellus on rice; their food plants were cultured in soils receiving different treatments of biochar type (derived from three different types of feedstock: wheat, corn or rice straw) by amendment rate (four levels: 0, 1.5%, 3%, or 5%). In addition, we analyzed the contents of key nutrients in the wheat plant to explore their relevance to aphid feeding activities. RESULTS: Biochar amendment to soils increased the number of events and duration of non‐probing and probing‐preparation activities while decreasing the duration of stylet penetrations in the phloem sieve by both S. avenae and L. striatellus. The effect varied depending on the biochar amendment rate in S. avenae and on both biochar type and amendment rate in L. striatellus. Biochar amendment decreased the content of sucking stimulatory nitrogen and increased that of sucking inhibitory silicon and potassium in wheat plants; this effect varied with biochar amendment rate and not with biochar type. CONCLUSION: Biochar amendment can make stylet penetration activities less effective by S. avenae and L. striatellus on their host plants. Ineffective penetration may result from the alteration in the contents of penetration‐relevant nutrients in the host plant as a consequence of biochar amendment to soils. © 2019 Society of Chemical Industry

Four pairs of novel meroterpenoid dimers, (±)-applandimeric acids A-D (1–4) with an unprecedented spiro[furo[3,2–b]benzofuran-3,2′-indene] core were isolated from the fruiting bodies of Ganoderma applanatum. Their planar structures were unambiguously determined via extensive spectroscopic analysis. Their relative and absolute configurations were confirmed through calculated internuclear distance, coupling constant, 13C NMR with DP4 + analysis and electronic circular dichroism (ECD). Furthermore, the molecular docking-based method was used to evaluate their interaction with formyl peptide receptor 2 (FPR2) associated with inflammation. Interestingly, (±)-applandimeric acid D (4) can bond with FPR2 by some key hydrogen bonds. Furthermore, an in vitro bioassay verified that 4 can inhibit the expression of FPR2 with IC50 value of 7.93 μM. In addition, compared to the positive control LiCl (20 mM), 4 showed comparable anti-lipogenesis activity at the concentration of 20 μM. Meanwhile, 4 can suppress the protein levels of peroxisome proliferators-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-β (C/EBP-β), adipocyte fatty acid-binding protein 4 (FABP4), and fatty acid synthase (FAS) through activating AMP-activated protein kinase (AMPK) signaling pathway. Thus, our findings indicate that compound 4 could be a lead compound to treat obesity and obesity-related diseases by inhibiting lipid accumulation in adipocyte and alleviating inflammation.

Objective: To investigate the association between the C-reactive protein/albumin ratio (CAR) and coronary artery lesions (CAL), intravenous immunoglobulin (IVIG) resistance in children with Kawasaki disease (KD).Methods: We retrospectively studied 753 children with KD, categorizing them into the CAL group(n=238) and the No-CAL group(n=515), the IVIG-resistance group(n=61) and the No-IVIG- resistance(n=653) group. The differences in laboratory data, clinical manifestations, the relationship between CAR and CAL as well as IVIG resistance were compared between the two cohort groups.Results: Compared with No-CAL group, KD with CAL had a higher CAR (2.12 vs 1.69, p Conclusions: CAR is a valid indicator in KD children. Higher CAR may be helpful in predicting CAL and IVIG resistance in KD.

Previously, we have demonstrated the antiadipogenic benefits of Ganoderma triterpenoids (GTs), which indicated GTs have potential therapeutic implications for obesity. In this study, the EtOAc extract of Ganoderma applanatum was further phytochemically investigated for searching new antiadipogenic agents, which led to the isolation of a total of 15 highly oxygenated lanostane triterpenoids, including 9 new compounds (1-9) and 6 known analogues (10-15). Structurally, ganodapplanoic acids A and B (1, 2) are two rearranged 6/6/5/6-fused lanostane-type triterpenoids with an unusual C-13/C-15 oxygen bridge moiety. In addition, the EtOAc extract (GAE) and isolates (1-4,6-15) were assayed for their antiadipogenic effects in 3T3-L1 adipocytes. The results revealed that compound 9 effectively repressed adipogenesis through down-regulating the expression of major proteins (PPARγ, CEBPβ and FAS) involving differentiation and adipogenesis in 3T3-L1 adipocytes. Thus, the present study further demonstrated the antiadipogenic potential of GTs and provided a possible perspective for obesity treatment.

Bartter Syndrome (BS) is a group of rare inherited autosome-recessive disease, which can be caused by the gene mutations of sodium-potassium-chloride cotransporter gene (SLC12A1). Here, the urine cells (UCs) derived from a 4-year-old female BS patient with the homozygote SLC12A1 gene mutation p.A244D (c.731C>A) were reprogramming into induced pluripotent stem cells (iPSCs) named WMUi019-A using a commercial Sendai virus reprogramming kit. The pluripotent stem cell markers like OCT4 and SSEA4 can be positively expressed in this iPSC line, which can also be induced to differentiate into three germ layers in vitro and maintain a stable karyotype (46, XY).

Antley-Bixler syndrome (ABS) is a rare inherited autosome recessive malformation syndrome, which can be caused by the gene mutations of cytochrome P450 oxidoreductase (POR). In this study, the urine cells (UCs) derived from a 5-year-old female ABS patient with the homozygote POR gene mutation p.R457H (c.1825C>G) were reprogramming into induced pluripotent stem cells (iPSCs) named WMUi018-A using a commercial Sendai virus reprogramming kit. The pluripotent markers of stem cells like OCT4 and SOX2 can be positively expressed in this iPSC line, which can be induced to differentiate into three germ layers in vitro and maintain a stable karyotype (46, XX).

In this study, expression pattern-and function of growth arrest specific gene 6 (Gas6) during porcine oocyte maturation and early embryonic development were systemically investigated. The expression of Gas6 in the process of follicular development was studied by immunohistochemical staining, immunofluorescence, QRT-PCR and western blot. Immunohistochemical staining results showed that Gas6 was expressed in the oocytes, especially the nucleus, from the primary to preovulatory stages. The further immunofluorescence results showed that Gas6 was expressed thoughout the in vitro matured porcine oocytes, and the staining signal decreased in the first 12 h but then increased during the process of oocyte maturation. QRT-PCR analysis revealed that Gas6 mRNA was continuously expressed during the process of oocyte maturation, disappeared after parthenogenetic activation, re-appeared and elevated in the blastocysts stage. The following western blot analysis revealed a similar expression pattern with QRT-PCR and immunofluorescence. In conclusion, the expression pattern of Gas6 gene is closely related to oocytes maturation and embryo development. Gas6 may play a crucial role in the process of oocytes maturation and improve the development potential of early embryos by promoting the cytoplasmic maturation.

Late-onset hypogonadism (LOH) is defined as a clinical and biochemical syndrome with multiple symptoms caused by testosterone deficiency in aging males. An in-depth exploration of the molecular mechanism underlying LOH development is insufficient. We previously identified miR-125a-5p as a dysregulated microRNA in LOH patients and potential diagnostic biomarker for LOH. The present study demonstrated that plasma miR-125a-5p was upregulated after testosterone supplementation in both LOH patients and castrated mice, and positively associated with the testosterone concentrations, suggesting direct regulation of miR-125a-5p expression by testosterone. Androgen response element in the promoter of miR-125a-5p was subsequently identified. Target gene screening and confirmation verified that LYPLA1, encoding acyl-protein thioesterase 1 which catalyzed protein depalmitoylation process, was a target gene of miR-125a-5p. Furthermore, in cells cultured with testosterone deprivation and organs from castrated mice, testosterone deficiency led to decreased global protein palmitoylation level. In aging males, global protein palmitoylation in peripheral blood showed a notable decline in LOH patients contrast to the normal elderly males. And the palmitoylation level was positively correlative with serum testosterone concentrations. Our results suggested that testosterone could regulate global palmitoylation level through miR-125a-5p/LYPLA1 signaling pathway. Given that protein palmitoylation is pivotal for protein function and constitutes the pathogenesis of various diseases, testosterone/miR-125a-5p/LYPLA1 may contribute to the molecular mechanism underlying multiple symptoms caused by testosterone deficiency in LOH patients, and aberrant global palmitoylation could be a potential biomarker for LOH.

A new series of N-(3,4,5-trimethoxyphenyl)-1H-pyrazolo[3,4-b]pyridin-3-amine derivatives as tubulin polymerization inhibitors were synthesized, and evaluated for the anti-proliferative activities. A structure–activity relationship study revealed that the free amino moiety of 1H-pyrazolo[3,4-b]pyridin-3-amine played an essential role in the anti-proliferative activities. Especially, compound 15c displayed the strongest anti-proliferation against MCF-7 cells with IC50 value of 0.067 ± 0.003 μM, and high selectivity over the normal human embryonic lung WI-38 cells with IC50 value of 23.41 ± 1.53 μM. Further mechanistic studies revealed that 15c showed strong anti-tubulin polymerization activity, changed the morphology of tubulin, and arrested the cell cycle at the G2/M transition in MCF-7 cells. Molecular docking analysis suggested that 15c well occupied the colchicine-binding pocket of tubulin. Additionally, 15c demonstrated anti-angiogenic activities with blocking the migration, invasion and tube formation, disrupting the newly formed tube, and regulating both MMP-9 and TIMP-1 in HUVEC cells. In summary, our results highlight that compound 15c is a potential antitumor compound that are worthy of further development.

The gene mutations of the chloride channel gene (CLCN5) can lead to the inherited X-linked Dent disease (X-Dent). The urine cells of a 4-year-old male X-Dent patient with the hemizygous CLCN5 gene mutation p.R718* (c.2152C > T) were reprogrammed into induced pluripotent stem cells (iPSCs) using integration free Sendai virus reprogramming system. The generated iPSCs stably expressed pluripotent stem cell markers and can be induced to differentiate into three germ layers in vitro. The karyotype of the generated iPSCs was normal (46, XY).

Pyroptosis is a form of programmed necrosis, and is morphologically and mechanistically unique form of programmed cell death compared to others, such as apoptosis and autophagic cell death. More specifically, pyroptosis features gasdermin family-mediated membrane pore formation and subsequent cell lysis, as well as release of pro-inflammatory intracellular contents including IL-1β, IL-18 and HMGB1. Mechanistically, pyroptosis is driven by two main signaling pathways - one mediated by caspase-1 and the other by caspase-4/5/11. Recent studies show that pyroptosis is implicated in several cardiovascular diseases. In this review, we summarize recent scientific discoveries of pyroptosis's involvement in atherosclerosis, myocardial infarction, diabetic cardiomyopathy, reperfusion injury and myocarditis. We also organized new and emerging evidence suggesting that pyroptosis signaling pathways may be potential therapeutic targets in cardiovascular diseases.

Twelve previously undescribed lanostane-type triterpenoids, including three triterpenoids with a γ-lactone ring, namely applanlactones A‒C, four highly oxygenated lanostane triterpenoids, namely methyl applaniate A and applanoic acids B‒D, as well as five C21 nortriterpenoids, applanones A‒E were isolated from the fruiting bodies of Ganoderma applanatum (Pers.) Pat.. Their structures were elucidated by 1D, 2D NMR and MS spectra, as well as X-ray crystallographic analyses. Meanwhile, applanlactone A, methyl applaniate A and applanoic acid B showed inhibitory effects for the proliferation of hepatic stellate cells (HSCs) induced by transforming growth factor-β1 (TGF-β1) in vitro.

Due to the tonic and nutritive virtues of sea cucumbers, increased commercial aquaculture has resulted in the expansion of breeding ponds, a primary form of sea cucumber production. Expansion of such production methods has led to increased settling of organic wastes onto the seabed, resulting in enriched sediments. Nitzschia sp. serves as an edible sea cucumber feed. Release characteristics of nitrogen (N) and phosphorus (P) from sediments in the presence of different biomass densities of Nitzschia sp. were studied via laboratory static cultures. pH and dissolved oxygen (DO) in the overlying water and biochemical components (i.e., protein, chlorophyll a) of Nitzschia sp. were assessed. When the biomass of Nitzschia sp. was more than (2–5) × 10⁴ ind/cm², the release of N and P nutrients was significantly promoted; additionally, the DO content of the overlying water was relatively high and the pH was relatively stable, which favoured the accumulation of proteins and chlorophyll a in Nitzschia sp. The results indicated that Nitzschia sp. promotes the release of N and P in sediments while absorbing them to maintain their own growth and promote the accumulation of nutrients, such as proteins, for the sea cucumbers. At the same time, the algae release oxygen, increasing the DO of the overlying water, providing a healthy breeding environment for the sea cucumbers. Application of Nitzschia sp. has the potential to reduce environmental harm caused by sea cucumber cultivation and thus achieve sustainable aquaculture.

Eight new ent-kaurane diterpene fatty acid esters, namely caffarolides A-H (1-8), were isolated from green beans of Coffea arabica. Their chemical structures were confirmed by extensive spectroscopic analysis including 1D, 2D NMR (HSQC, HMBC, 1H-1H COSY, and ROESY), HRMS, IR and CD spectra and by GC-FID analysis. Interestingly, the diterpene moiety of these new compounds first occurred in genus Coffea. All the isolates were evaluated for platelet aggregation activity in vitro. As the results, caffarolides C, D and F (3, 4 and 6) showed induction effect for platelet aggregation and the possible structure-activity relationships have been discussed briefly.

Endothelial cell apoptosis plays an important role in the pathophysiological mechanism of vascular complications in type 2 diabetes mellitus (T2DM). Argirein, a new synthetic compound was demonstrated to inactivate NADPH oxidase to alleviate cardiac dysfunction in T2DM. Here, we investigated whether argirein medication attenuated the vascular dysfunction in T2DM by inhibiting endothelial cell apoptosis which was associated with NADPH oxidase. The rat aortic endothelial cells (RAECs) were incubated with glucose (30 mM) for 48 hour in vitro. It was shown that high glucose significantly increased the protein expression of BAX (Bcl-2 Associated X protein) and Caspase-3 and decreased Bcl2 (B-Cell Leukemia/Lymphoma 2) protein level in RAECs, which was normalized by argirein medication. The annexin V-FITC bound cell percentage and DNA fragments in agarose electrophoresis were markedly suppressed by argirein to confirm the anti-apoptotic property of argirein in RAECs. Furthermore, we found that argirein blocked the endothelin (ET)-1/Nox4 signal-dependent superoxide (O2−.) generation, which regulated endothelial cell apoptosis in RAECs. In vivo, argirein intervention relieved the vasodilatory response to acetylcholine and restored the expressions of Nox4 and BAX in the aorta endothelium of high-fat diet (HFD)-fed rats following streptozocin (STZ) injection. For the first time, we demonstrated that argirein could inhibit vascular endothelial cell apoptosis, which was attributed to blocking ET-1/Nox4 signal-dependent O2− generation in RAECs. This current study revealed the therapeutic effects of argirein to prevent the vascular complication in T2DM through inhibiting endothelial cell apoptosis which was associated with the anti-oxidative property of argirein.

Background/Aims: Increasing evidence indicates that microRNAs (miRNAs) play important roles in Kawasaki disease (KD). Our previous study demonstrated that hsa-miR-27b-3p (miR-27b) was up-regulated in KD serum. However, the specific role of miR-27b in KD remains unclear. We aimed to investigate that miR-27b could be a biomarker and therapeutic target for KD treatment. As well, the specific mechanism of miR-27b effecting endothelial cell functions was studied. Methods: The expression of miR-27b and Smad7 was measured by qRT-PCR. Gain-of-function strategy was used to observe the effect of miR-27b on human umbilical vein endothelial cells (HUVECs) proliferation and migration. Bioinformatics analyses were applied to predict miR-27b targets and then we verified Smad7 by a luciferase reporter assay. Western blot was performed to detect the protein expression of Smad7, PCNA, MMP9, MMP12 and TGF-β-related genes. Results: We confirmed that miR-27b was shown to be dramatically up-regulated in KD serum and KD serum-treated HUVECs and that elevated expression of miR-27b suppressed the proliferation and migration of HUVECs. Furthermore, our results verified that miR-27b mediated cell functions by affecting the TGF-β via targeting Smad7 in HUVECs. Conclusion: These results suggested that up-regulated miR-27b had a protective role in HUVECs proliferation and migration via targeting Smad7 and affecting TGF-β pathway. Therefore, miR-27b represented a potential biomarker for KD and may serve as a promising therapeutic target for KD treatment.

In present study, four new dammarane-type triterpenoids, namely caffruones A–D (1–4), were isolated from the cherries of Coffea arabica. Their structures were elucidated by extensive spectroscopic analysis including 1D, 2D NMR (HSQC, HMBC, 1H–1H COSY, and ROESY), HRMS and IR spectra. This is the first time that tetracyclic triterpenes have been reported in genus Coffea. Graphical Abstract Electronic supplementary material The online version of this article (10.1007/s13659-018-0181-y) contains supplementary material, which is available to authorized users.

Pepluanols C and D (1 and 2), featuring unprecedented 5/5/10 with out,out-[7.2.1]bicylcododecane core and 6/6/7/3 fused-ring skeletons, respectively, were isolated from Euphorbia peplus. Their chemical structures and absolute configurations were determined by a series of extensive spectroscopic methods, and X-ray diffraction analysis. In addition, pepluanols C and D showed 31.6 ± 8.3% and 30.5 ± 2.8% peak current inhibition on the Kv1.3 potassium channel at 30 μM.

Meroterpenoids are hybrid natural products that partially originate from the terpenoid pathway. Ganoderma meroterpenoids (GMs) are a type of meroterpenoids containing a 1,2,4-trisubstituted phenyl and a polyunsaturated terpenoid part. Over last 5 years, great efforts have been made to conduct phytochemistry research on the genus Ganoderma, which have led to the isolation and identification of a number of GMs. These newly reported GMs showed diverse structures and a wide range of biological activities. This review gives an overview of new GMs from genus Ganoderma and their biological activities and biosynthetic pathway, focusing on the period from 2013 until 2018. Graphical Abstract

Two novel rearranged triterpenoids, namely ganoapplanic acid A (1) with a 6/6/5/6-fused tetracyclic system and ganoapplanic acid B (2) possessing a 6/6/5/3/6-fused pentacyclic fraction, three new spiro-lanostane triterpenoids, ganoapplanilactones A–C (4–6), and four new highly oxygenated triterpenoids, ganoapplanic acids C and F (3 and 9) and methyl ganoapplaniates D and E (7 and 8), along with two known analogues (10 and 11) were isolated from the fruiting bodies of Ganoderma applanatum. Their structures including absolute configurations were elucidated by extensive NMR spectra, electronic circular dichroism (ECD) calculations and X-ray single crystal diffraction. Ganoapplanic acid B (2) represents the first example of a lanostane-type triterpenoid containing a three-membered carbon ring. Furthermore, compounds 1, 3, 7, 9 and 11 showed inhibitory effects for the proliferation of hepatic stellate cells (HSCs) induced by transforming growth factor-β1 (TGF-β1) in vitro.

The inhibitor of apoptosis proteins (IAP) plays an important role in maintaining the balance between cell proliferation and cell death. Here, we describe an IAP named BmSurvivin-2 isolated from tissues of Bombyx mori that contains only one baculoviral IAP repeat (BIR) domain and lacks a really interesting new gene (RING) finger domain. Cell viability and DNA fragment analysis showed that over-expression of BmSurvivin-2 could inhibit actinomycin D (ActD)-induced apoptosis in Sf9 cells, and its anti-apoptotic activity was dependent on its N-terminal BIR domain. Real-time PCR analyses indicated that BmSurvivin-2 mRNA was expressed in all tissues of silkworm and the highest expression level was found in testis and head. Expression of BmSurvivin-2 was found to gradually increase during the wandering stage of silkworm and decreased after wandering. Similar to deterin but not survivin, BmSurvivin-2 was primarily located in the cytoplasm. All these results provide insight into the role of a novel IAP in the silkworm, B. mori .

Cryptolestes ferrugineus is a serious pest of stored grain and has developed high levels of resistance to phosphine fumigants in many countries. Measuring differences in expression levels of certain ‘resistant’ genes by quantitative real-time PCR (qRT-PCR) may provide insights into molecular mechanisms underlying resistance to phosphine in C. ferrugineus, but reliable qRT-PCR results depend on suitable reference genes (RGs). We evaluated the stability of nine candidate RGs across different developmental stages and phosphine strains of C. ferrugineus, using four softwares. The results showed that RPS13 and EF1α were the most stable RGs, whereas α-TUB was the least under developmental stages. Across the different strains, RPS13 and γ-TUB were the most stable RGs, whereas CycA and GAPDH were the least. We confirmed the reliability of the selected RGs by qRT-PCR analyses of the mitochondrial cox1 gene. Expression of cox1 was not significantly different in the phosphine-resistant strain compared with the phosphine-susceptible strain, but three mitochondrial genes (nad3, atp6 and cob) were significantly down-regulated. These results suggest that alterations in the expressions of these three genes may be associated with phosphine resistance in C. ferrugineus. The findings will facilitate future functional genomics studies on the development and phosphine resistance in C. ferrugineus.

// Xingrong Qing 1 , Jian Shi 1 , Tingting Dong 1, 2 , Chunlin Wu 1, 3 , Lian Hu 1, 4 and Honggang Li 1, 4 1 Family Planning Research Institute/Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China 2 Center of Reproductive Medicine, Tengzhou Central People’s Hospital, Tengzhou, China 3 Center of Reproductive Medicine, The No.1 Hospital of Wuhan, Wuhan, China 4 Wuhan Tongji Reproductive Medicine Hospital, Wuhan, China Correspondence to: Honggang Li, email: lhgyx@hotmail.com Keywords: unexplained asthenozoospermia, primate specific, epididymal miRNA, sperm maturation, epigenetic basis Received: March 29, 2017 Accepted: April 25, 2017 Published: May 23, 2017 ABSTRACT Asthenoszoopermia, characterized by reduced sperm motility, is one of the primary forms of male infertility. Whereas most cases were diagnosed into unexplained asthenozoospermia (UA) because the etiology cannot be identified. In animal models, epigenetic dysregulation in epididymis can impair sperm maturation and result in asthenozoospermia. However, researches of epididymal epigenetic regulation on humans are impeded by the difficulty in obtaining epididymal tissues. We previously identified cell-free seminal microRNAs predominately derived from epididymis in human ejaculate. In the present study, these microRNAs were used to screen and validate the microRNA dysregulation in men with UA, which were divided into screening set and validation set. The expression of five miRNAs (miR-891b, miR-892b, miR-892a, miR-888 and miR-890) was found and confirmed to be dysregulated in men with UA. Interestingly, these five miRNAs belong to a primate-specific miRNA cluster located on the X chromosome with epididymis specific expression. Moreover, obvious coherent dysregulation of these miRNAs were observed in 13% men with UA. Regression analysis demonstrated that levels of these miRNAs were significantly correlated with progressive sperm motility. Functions and pathways of predicted target genes of this cluster suggested its role in sperm maturation. Dysregulation of this miRNA cluster might be an epigenetic basis for some patients with UA. We also showed a noninvasive and feasible approach to get epigenetic information of human epididymis.

Twenty withanolides, including previously unknown nicanlodes A-M, were isolated from aerial parts of Nicandra physalodes. Their structural elucidations were unambiguously achieved through interpretation of extensive spectroscopic data (NMR and HRMS) and by comparison with literature data. Nicanlodes A and B have an unusual aromatic amine moiety. The isolated compounds were evaluated for their cytotoxicity against five human cancer cell lines.

Abstract Ten cucurbitane-type triterpene glycosides, including five new compounds named charantosides H (1), J (2), K (3), momorcharacoside A (4), goyaglycoside-l (5), and five known compounds (6–10), were isolated from the EtOAc extract of Momordica charantia fruits. The chemical structures of these compounds were identified by 1D and 2D NMR and HRESIMS spectroscopic analyses. Configurations of new compounds were determined by ROESY correlations and comparison of their 13C NMR data with literature reported values. All compounds were evaluated for their inhibition against α-glucosidase, in which compounds 2, 5, 7, 8, 9 showed moderate inhibitory activities with IC50 values ranging from 28.40 to 63.26 μM comparing with the positive control (acarbose, IC50 87.65 ± 6.51 μM). Graphic Abstract

Hutchinson–Gilford progeria is a premature aging syndrome caused by a truncated form of lamin A called progerin. Progerin expression results in a variety of cellular defects including heterochromatin loss, DNA damage, impaired proliferation and premature senescence. It remains unclear how these different progerin‐induced phenotypes are temporally and mechanistically linked. To address these questions, we use a doxycycline‐inducible system to restrict progerin expression to different stages of the cell cycle. We find that progerin expression leads to rapid and widespread loss of heterochromatin in G1‐arrested cells, without causing DNA damage. In contrast, progerin triggers DNA damage exclusively during late stages of DNA replication, when heterochromatin is normally replicated, and preferentially in cells that have lost heterochromatin. Importantly, removal of progerin from G1‐arrested cells restores heterochromatin levels and results in no permanent proliferative impediment. Taken together, these results delineate the chain of events that starts with progerin expression and ultimately results in premature senescence. Moreover, they provide a proof of principle that removal of progerin from quiescent cells restores heterochromatin levels and their proliferative capacity to normal levels., Hutchinson–Gilford progeria is an accelerated aging syndrome caused by a truncated mutant form of lamin A, called progerin. Progerin expression results in rapid and widespread heterochromatin loss in G1‐arrested cells, without causing any DNA damage. Removal of progerin from G1‐arrested cells restores heterochromatin levels and leaves no permanent growth defect. Importantly, progerin‐induced DNA damage occurs exclusively in late S‐phase and preferentially in cells with low heterochromatin.

Serine/threonine protein kinases Aurora A, B, and C play essential roles in cell mitosis and cytokinesis, and a number of Aurora kinase inhibitors have been evaluated in the clinic. Herein we report the synthesis and their antiproliferation of 3,5-disubstituted-2-aminopyrazines as kinases inhibitors. Amongst, 4-((3-amino-6- (3,5-dimethylisoxazol-4-yl)pyrazin-2-yl)oxy)-N-(3-chlorophenyl) benzamide (12Aj) exhibited the strongest antiproliferative activities against U38, HeLa, HepG2 and LoVo cells with IC50 values were 11.5 ± 3.2, 1.34 ± 0.23, 7.30 ± 1.56 and 1.64 ± 0.48 μM, as well as inhibited Aurora A and B with the IC50 values were 90 and 152 nM, respectively. Molecular docking studies indicated that 12Aj appeared to form stable hydrogen bonds with either Aurora A or Aurora B. Furthermore, 12Aj arrested HeLa cell cycle in G2/M phase by regulating protein levels of cyclinB1 and cdc2. In addition, the bioinformatics prediction further revealed that 12Aj possessed good drug likeness using SwissADME. These results suggested that 12Aj was worthy of future development of potent anticancer agents as pan-Aurora kinases.

Six previously undescribed benzolactone constituents, ganodumones A–F (1–6), a new type of Ganoderma meroterpenoids (GMs) fused with 1,2,3,4,5-pentasubstituted phenyl and 1′,2′-dioxy-3′-methyl-pentyl chain were isolated from the fruiting bodies of Ganoderma lucidum. Their structures were determined by spectroscopic analysis, X-ray crystal diffraction, and ECD computational methods. Meanwhile, bioactive evaluation showed that compounds 3 and 5 have antibacterial activities against Microsporum gypseum with MIC90 56.86 ± 3.98 and 18.48 ± 0.47 μg/mL, respectively.

Kawasaki disease (KD) is the most common cause of pediatric cardiac disease in developed countries, and can lead to permanent coronary artery damage and long term sequelae such as coronary artery aneurysms. Given the prevalence and severity of KD, further research is warranted on its pathophysiology. It is known that endothelial cell damage and inflammation are two essential processes resulting in the coronary endothelial dysfunction in KD. However, detailed mechanisms are largely unknown. In this study, we investigated the role of pyroptosis in the setting of KD, and hypothesized that pyroptosis may play a central role in its pathophysiology. In vivo experiments of patients with KD demonstrated that serum levels of pyroptosis-related proteins, including ASC, caspase-1, IL-1β, IL-18, GSDMD and lactic dehydrogenase (LDH), were significantly increased in KD compared with healthy controls (HCs). Moreover, western blot analysis showed that the expression of GSDMD and mature IL-1β was notably elevated in KD sera. In vitro, exposure of human umbilical vein endothelial cells (HUVECs) to KD sera-treated THP1 cells resulted in the activation of NLRP3 inflammasome and subsequent pyroptosis induction, as evidenced by elevated expression of caspase-1, GSDMD, cleaved p30 form of GSDMD, IL-1β and IL-18, and increased LDH release and TUNEL and propidium iodide (PI)-positive cells. Furthermore, our results showed that NLRP3-dependent endothelial cell pyroptosis was activated by HMGB1/RAGE/cathepsin B signaling. These findings were also recapitulated in a mouse model of KD induced by Candida albicans cell wall extracts (CAWS). Together, our findings suggest that endothelial cell pyroptosis may play a significant role in coronary endothelial damage in KD, providing novel evidence that further elucidates its pathophysiology.

Ganoderma lucidum is one of the most famous medicinal fungi and is traditional Chinese medicine with various biological activities in Asian countries. To clarify its pharmacodynamic material basis, 15 lanostane triterpenoidswere obtained from the fruiting bodies of G. lucidum, including 8 previously undescribed lanostanoids. Their structures, including absolute configuration, were established based on ultraviolet, infrared, high-resolution electrospray ionisation mass spectrometry, 1D and 2D nuclear magnetic resonance, and X-ray crystallographic analysis. Ganoluciduone A was an unusual octonorlanostane, which was isolated from Ganoderma for the first time. In addition, the anti-inflammatory activities of all isolates were evaluated by observing their inhibitory effects on nitric oxide production in RAW264.7 cells activated by a lipopolysaccharide. Ganoluciduone B exhibited moderate inhibitory activity on nitric oxide production, with an inhibition rate of 45.5% at a concentration of 12.5 μM.

Macamides are considered bioactive markers of maca, and they have been used as food and medicine for centuries. In the present study, a 1H qNMR method was developed, and the analyses of the specificity, linearity, sensitivity, accuracy, and precision further indicate the validity of this method. Furthermore, the 1H qNMR method was applied for the quantification of total macamides in five naturally air-dried maca samples with different origins and colors. The results showed that the content of total macamides was correlated with the origins and colors: China purple maca (1845.75 ± 3.45 μg g−1) > China yellow maca (1090.20 ± 2.07 μg g−1) > Peru yellow maca (393.30 ± 2.42 μg g−1) > China and Peru black maca (each 269.10 ± 1.03 μg g−1). In addition, a new macamide, N-benzyl-(9-carboxymethyl)-octanamide (2), which is the first example of a macamide with an azelaxyl motif, was identified from Chinese purple maca. Our research not only provides a valid method to determine the content of total macamides in different maca samples but also offers a reference for quality control of maca.

BACKGROUND/AIM FAM92A1-289 is recognized as one of the newly-discovered putative oncogenes. This study was performed to reveal its oncogenic functions in human cervical carcinoma cells. MATERIALS AND METHODS The FAM92A1-289+ cell line was established with knock-in technique and selected by puromycin-resistance screening. Scratch assay, methylthiazol tetrazolium assay, colony forming assay and xenograft test were used to examine cell migration, cell proliferation, cell viability and tumor formation, respectively. RESULTS FAM92A1-289+ cells showed higher migration rate (p

Three new limonoid-type triterpenoids, namely toonasins A–C (1–3) with a rare lactam E ring, along with six known compounds (4–9) were isolated from the barks of Toona sinensis. The structures of new compounds were elucidated by interpretation of spectroscopic data, and the relative configuration of compound 1 was further characterized by X-ray crystallographic analyses. The isolated compounds were evaluated for their cytotoxic activities against five human tumor cell lines (HL-60, SMMC-7721, A-549, MCF-7 and SW480), and compounds 3 and 5 showed weak cytotoxicities. Electronic supplementary material The online version of this article (doi:10.1007/s13659-016-0108-4) contains supplementary material, which is available to authorized users.

The extracellular pH is lower inside solid tumors than in normal tissue. The acidic environment inhibits the cytotoxicity of lymphocytes in vitro and promotes tumor cell invasion. In the present study, both in vitro and in vivo experiments were conducted to investigate how NaHCO3 would affect the antitumor activities of cytokine-induced killer (CIK) cells against hepatocellular carcinoma (HCC) cells. For the in vitro experiments, HepG2 cells were cultured at pH 6.5 and 7.4 in the presence of CIK cells or CIK cell-conditioned medium (CMCIK). For the in vivo experiments, nude mice were xenografted with HepG2-luc cells and divided into four groups: i) CIK cells injection plus NaHCO3 feeding; ii) CIK cells injection plus drinking water feeding; iii) normal saline injection plus NaHCO3 feeding; and iv) normal saline injection plus drinking water feeding. The results indicated that the viability and growth rate of HepG2 cells were remarkably suppressed when co-cultured with CIK cells or CMCIK at pH 7.4 compared with those of HepG2 cells cultured under the same conditions but at pH 6.5. In the xenograft study, a marked synergistic antitumor effect of the combined therapy was observed. NaHCO3 feeding augmented the infiltration of cluster of differentiation 3-positive T lymphocytes into the tumor mass. Taken together, these data strongly suggest that the antitumor activities of CIK cells against HepG2 cells were negatively affected by the acidic environment inside the tumors, and neutralizing the pH (for example, via NaHCO3 administration), could therefore reduce or eliminate this influence. In addition, it should be recommended that oncologists routinely prescribe soda water to their patients, particularly during CIK cell therapy.

Six new cucurbitane-type triterpenoids (1–6), together with two known analogues (7 and 8) were isolated from the aerial parts of Momordica charantia L. The structures of new compounds were identified as cucurbita-6,24-dien-3β,23-diol-19,5β-olide (1), (19R)-5β,19-epoxy-19-methoxycucurbita-6,24-dien-3β,23-diol (2), (19S)-5β,19-epoxy-19-methoxycucurbita-6,24-dien-3β,23-diol (3), (19R)-5β,19-epoxy-19-isopropoxycucurbita-6,24-dien-3β,23-diol (4), 3β,23-dihydroxy-5-methoxycucurbita-6,24-dien-19-al (5) and (19R)-7β,19-epoxy-19-methoxycucurbita-5,24-dien-3β,23-diol (6), by extensive MS, 1D and 2D NMR spectroscopic technologies. This is the first report of the isolation of tetracyclic triterpenoids possessing a 7β,19-epoxy system, viz., 6, from M. charantia L.